2022/04/12 更新

写真a

アサヌマ ヒロユキ
浅沼 浩之
ASANUMA, Hiroyuki
所属
大学院工学研究科 生命分子工学専攻 分子生命化学 教授
大学院担当
大学院工学研究科
学部担当
工学部 化学生命工学科
職名
教授
連絡先
メールアドレス

学位 1

  1. 工学博士 ( 1989年3月   東京大学 ) 

研究キーワード 3

  1. 核酸化学

  2. 光化学

  3. 高分子化学

研究分野 1

  1. その他 / その他  / 生体関連化学

現在の研究課題とSDGs 6

  1. 核酸機能の光制御を目指した光応答性DNA/RNAの開発

  2. DNA/RNAを認識する高感度蛍光プローブの開発

  3. DNA/RNAを足場とする新規色素クラスターの設計

  4. 非環状型人工核酸SNA, aTNAの開発

  5. 核酸医薬への応用を目指した人工核酸の開発

  6. 人工核酸の非酵素的複製の研究

▼全件表示

経歴 4

  1. 名古屋大学   大学院工学研究科 生命分子工学専攻   教授

    2022年4月 - 現在

  2. 名古屋大学   予防早期医療創成センター   教授

    2020年5月 - 2022年3月

  3. 名古屋大学   大学院工学研究科生命分子工学専攻   教授

    2017年4月 - 2020年5月

  4. 名古屋大学   大学院工学研究科 生命分子工学専攻 分子生命化学   教授

    2017年4月 - 2020年4月

学歴 3

  1. 東京大学   大学院工学系研究科   工業化学専門課程博士課程

    1986年4月 - 1989年3月

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    国名: 日本国

  2. 東京大学   大学院工学系研究科   工業化学専門課程修士課程

    1984年4月 - 1986年3月

  3. 東京大学   工学部   合成化学科

    1980年4月 - 1984年3月

所属学協会 6

  1. 生命の起源および進化学会   会員

    2021年4月 - 現在

  2. 日本核酸化学会   運営委員

    2017年9月 - 現在

  3. 高分子学会

    2011年4月 - 現在

  4. 日本化学会

  5. 光化学協会   会員

  6. 光化学協会

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委員歴 8

  1. 日本核酸化学会   運営委員  

    2017年9月 - 現在   

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    団体区分:学協会

  2. Wiley-VCH   Editorial Advisory Board ChemBioChem  

    2016年1月 - 現在   

  3. 日本化学会生体機能関連化学・バイオテクノロジーディビジョン   幹事  

    2014年9月 - 現在   

  4. 日本化学会生体機能関連化学・バイオテクノロジーディビジョン   幹事  

    2014年9月 - 現在   

  5. 生体機能関連化学部会   部会長  

    2012年4月 - 現在   

  6. 国際核酸化学シンポジウム実行委員会   実行委員  

    2011年9月 - 2012年8月   

  7. 高分子学会東海支部   常任幹事  

    2010年4月 - 現在   

  8. Associate editor of Bulletin of the Chemical Society of Japan  

    2006年4月 - 2010年3月   

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    団体区分:学協会

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受賞 4

  1. 平成29年度日本化学会 学術賞

    2018年3月   公益社団法人日本化学会   非環状型核酸アナログによる機能性オリゴヌクレオチドの創製

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    受賞区分:国内学会・会議・シンポジウム等の賞  受賞国:日本国

  2. 平成27年度高分子学会賞(科学)

    2016年5月   公益社団法人高分子学会  

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    受賞国:日本国

  3. 部会講演賞

    2000年9月   日本化学会生体機能関連化学部会   アゾベンゼン導入DNAによる二重鎖形成の光制御機構の解析

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    受賞国:日本国

  4. 若い世代の特別講演会講演証

    1990年4月   日本化学会   乾燥高分子ゲル錯体の多孔性付与による高機能化

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    受賞区分:国内学会・会議・シンポジウム等の賞  受賞国:日本国

 

論文 231

  1. Nonenzymatic polymerase-like template-directed synthesis of acyclic L-threoninol nucleic acid 査読有り

    Murayama, K.; Okita, H.; Kuriki, T.; Asanuma, H.

    Nature Communications   12 巻   頁: 804   2021年2月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: https://doi.org/10.1038/s41467-021-21128-0

  2. Orthogonal Amplification Circuits Composed of Acyclic Nucleic Acids Enable RNA Detection 査読有り

    Chen Y., Nagao R., Murayama K., Asanuma H.

    Journal of the American Chemical Society     2022年

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Journal of the American Chemical Society  

    Construction of complex DNA circuits is difficult due to unintended hybridization and degradation by enzymes under biological conditions. We herein report a hybridization chain reaction (HCR) circuit composed of left-handed acyclic d-threoninol nucleic acid (d-aTNA), which is orthogonal to right-handed DNA and RNA. Because of its high thermal stability, use of an aTNA hairpin with a short 7 base-pair stem ensured clear ON-OFF control of the HCR circuit. The aTNA circuit was stable against nucleases. A circuit based on right-handed acyclic l-threoninol nucleic acid (l-aTNA) was also designed, and high orthogonality between d- and l-aTNA HCRs was confirmed by activation of each aTNA HCR via a corresponding input strand. A dual OR logic gate was successfully established using serinol nucleic acid (SNA), which could initiate both d- and l-aTNA circuits. The d-aTNA HCR was used for an RNA-dependent signal amplification system via the SNA interface. The design resulted in 80% yield of the cascade reaction in 3000 s without a significant leak. This work represents the first example of use of heterochiral HCR circuits for detection of RNA molecules. The method has potential for direct visualization of RNA in vivo and the FISH method.

    DOI: 10.1021/jacs.1c12659

    Scopus

  3. Color-Changing Fluorescent Barcode Based on Strand Displacement Reaction Enables Simple Multiplexed Labeling 査読有り

    Makino, K.; Susaki, E.; Endo, M.; Asanuma, H.; Kashida, H.

    Journal of the American Chemical Society   144 巻   頁: 1572 - 1579   2022年1月

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    担当区分:責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1021/jacs.1c09844

  4. Xeno nucleic acids (XNAs) having non-ribose scaffolds with unique supramolecular properties 招待有り 査読有り

    Asanuma, H.; Kamiya, Y.; Kashida, H.; Murayama, K.

      28 巻   頁: 3933   2022年3月

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1039/d1cc05868a

  5. Pseudo base pairs that exhibit high duplex stability and orthogonality through covalent and non─covalent interactions 招待有り

    樫田 啓, 浅沼 浩之

    有機合成化学協会誌   79 巻 ( 11 ) 頁: 1013 - 1019   2021年11月

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    担当区分:最終著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 有機合成化学協会  

    Natural base pairs stabilize nucleic acid duplexes via hydrogen bonding and stacking interactions, and efforts have been made to identify artificial base pairs that stabilize duplexes through different interactions. Herein, we describe our attempts to prepare pseudo base pairs that exhibit high stability and orthogonality. Pairs of azobenzene derivatives showed unexpectedly high stability and orthogonality due to intermolecular stacking interactions. Pairs of cationic molecules showed even higher stability than natural G–C pairs through both electrostatic and stacking interactions. Duplexes can be crosslinked via [2+2] photocycloaddition between stilbene derivatives, and pairs of non–planar cyclohexane derivatives can stabilize duplexes by hydrophobic interactions rather than stacking interactions. Hetero–selective pairing can be achieved using electron donor–acceptor interactions. These pseudo base pairs would work well as building blocks in nanotechnology and biotechnology due to their unique functionalities.

    DOI: 10.5059/yukigoseikyokaishi.79.1013

    Scopus

    CiNii Research

  6. Perylene-Cy3 FRET System to Analyze Photoactive DNA Structures 査読有り

    Kawai H., Doi T., Ito Y., Kameyama T., Torimoto T., Kashida H., Asanuma H.

    Chemistry - A European Journal   27 巻 ( 50 ) 頁: 12845 - 12850   2021年9月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Chemistry - A European Journal  

    We report a new Förster resonance energy transfer (FRET) system for structural analyses of DNA duplexes using perylene and Cy3 as donor and acceptor, respectively, linked at the termini of a DNA duplex via D-threoninol. Experimentally obtained FRET efficiencies were in good agreement with theoretical values calculated based on canonical B-form DNA. Due to the relatively long Förster radius, this system can be used to analyze large DNA structures, and duplexes containing photo-reactive molecules can be analyzed since perylene can be excited with visible light. The system was used to analyze a DNA duplex containing stilbene, demonstrating that in the region of the stilbene cluster the duplex adopts a ladder-like structure rather than helical one. Upon photodimerization between stilbene residues, FRET efficiencies indicated the reaction does not disturb DNA duplex. This FRET system will be useful for analysis of photoreactions of nucleobases as well as a wide range of nucleic acid structures.

    DOI: 10.1002/chem.202101738

    Scopus

    PubMed

  7. Design and Hybridization Properties of Acyclic Xeno Nucleic Acid Oligomers 招待有り 査読有り

    Murayama K., Asanuma H.

    ChemBioChem   22 巻 ( 15 ) 頁: 2507 - 2515   2021年8月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ChemBioChem  

    Xeno nucleic acids (XNAs) are analogues of DNA and RNA that have a non-ribose artificial scaffold. XNAs are possible prebiotic genetic carriers as well as alternative genetic systems in artificial life. In addition, XNA oligomers can be used as biological tools. Acyclic XNAs, which do not have cyclic scaffolds, are attractive due to facile their synthesis and remarkably high nuclease resistance. To maximize the performance of XNAs, a negatively charged backbone is preferable to provide sufficient water solubility; however, acyclic XNAs containing polyanionic backbones suffer from high entropy cost upon duplex formation, because of the high flexibility of the acyclic nature. Herein, we review the relationships between the structure and duplex hybridization properties of various acyclic XNA oligomers with polyanion backbones.

    DOI: 10.1002/cbic.202100184

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    PubMed

  8. Preparation of Artificial Hexaplex Composed of Non-Natural Nucleic Acid 招待有り 査読有り

    Makino K., Hattori Y., Kashida H., Asanuma H.

    Current Protocols   1 巻 ( 4 ) 頁: e106   2021年4月

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    記述言語:日本語   出版者・発行元:Current Protocols  

    This article describes synthetic procedures for acyclic D-threoninol nucleic acid tethering of bifacial nucleobases. Because these nucleobases have complementary hydrogen bonding sites on both sides, their oligomers can form a hexaplex. These hexaplexes are suitable for use as metal or pH sensors and as supramolecular motifs. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Synthesis of D-threoninol backbone. Basic Protocol 2: Synthesis of D-aTNA tethering cyanuric acid. Basic Protocol 3: Synthesis of D-aTNA tethering a 2-aminopyrimidine moiety. Basic Protocol 4: Synthesis of D-aTNA tethering a 2,4,6-triaminopyrimidine moiety. Basic Protocol 5: Synthesis of D-aTNA oligomer tethering cyanuric acid, 2-aminopyrimidine, or 2,4,6-triaminopyrimidine.

    DOI: 10.1002/cpz1.106

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    PubMed

  9. Dual Crosslinking Photo-Switches for Orthogonal Photo-Control of Hybridization Between Serinol Nucleic Acid and RNA 査読有り

    Yamano Y., Murayama K., Asanuma H.

    Chemistry - A European Journal   27 巻 ( 14 ) 頁: 4599 - 4604   2021年3月

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    担当区分:最終著者, 責任著者   記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Chemistry - A European Journal  

    Wavelength-selective photo-regulation by multiple chromophores responding to different wavelengths can expand the variation of photo-manipulating systems. Herein, we report the orthogonal photo-regulation of duplex formation between serinol nucleic acid (SNA) and RNA using light-induced crosslinking reactions mediated by a new photo-reactive nucleobase 8-naphthylvinyladenine (NVA) and previously described 8-pyrenylvinyladenine (PVA). An intrastrand crosslink was induced in an SNA strand containing two adjacent NVA residues by irradiation with 340–405 nm light; the crosslink was reversed by irradiation with ≤300 nm light. In an SNA strand with adjacent NVA and PVA residues, an intrastrand crosslink resulted from irradiation with 405–465 nm light that was reversed by irradiation with ≤340 nm light. Intrastrand photo-crosslinking caused severe destabilization of an SNA/RNA duplex, resulting in dissociation to single strands. Cycloreversion resulted in duplex formation. With these NVA/NVA and NVA/PVA photo-switches, four hybridization states of two SNA/RNA duplexes could be orthogonally photo-controlled by irradiation with a suitable wavelength of light.

    DOI: 10.1002/chem.202003528

    Web of Science

    Scopus

    PubMed

  10. Development and modification of pre-mirnas with a fret dye pair for the intracellular visualization of processing intermediates that are generated in cells 招待有り 査読有り

    Kamiya Y., Kamimoto H., Zhu H., Asanuma H.

    Sensors   21 巻 ( 5 ) 頁: 1 - 14   2021年3月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Sensors  

    microRNAs (miRNAs) are small non-coding ribonucleic acids (RNAs), which regulate gene expression via the RNA interference (RNAi) system. miRNAs have attracted enormous interest because of their biological significance and disease relationship. In cell systems, the generation of miRNA is regulated by multiple steps: the transfer of primary miRNA from the nucleus to the cy-tosol, the generation of the precursor-miRNA (pre-miRNA), the production of double-stranded RNA from pre-miRNA by the Dicer, the interaction with protein argonaute-2 (AGO2), and the sub-sequent release of one strand to form miRISC with AGO2. In this study, we attempt to visualize the intermediates that were generated in the miRNA-maturation step in the cells to acquire a detailed understanding of the maturation process of miRNA. To achieve this, we developed pre-miRNAs labeling with a Dicer-or AGO2-responsible fluorescence resonance energy transfer (FRET) dye pair. We observed that modifications with the dye at suitable positions did not interfere with the biological activities of pre-miRNAs. Further, imaging analyses employing these pre-miRNAs demon-strated that the processing of pre-miRNA promoted the accumulation of miRNA at the specific foci in the cytosol. The FRET-labeled pre-miRNA would further elucidate the mechanisms of the RNAi process and provide the basis for development of nucleic acid drugs working in the RNAi system.

    DOI: 10.3390/s21051785

    Scopus

    PubMed

  11. A helical amplification system composed of artificial nucleic acids 査読有り

    Kashida H., Nishikawa K., Shi W., Miyagawa T., Yamashita H., Abe M., Asanuma H.

    Chemical Science   12 巻 ( 5 ) 頁: 1656 - 1660   2021年2月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Chemical Science  

    Herein we report an amplification system of helical excess triggered by nucleic acid hybridization for the first time. It is usually impossible to prepare achiral nanostructures composed of nucleic acids because of their intrinsic chirality. We used serinol nucleic acid (SNA) oligomers for the preparation of achiral nanowires because SNA oligomers with symmetrical sequences are achiral. Nanowire formation was confirmed by atomic force microscopy and size exclusion chromatography. When a chiral nucleic acid with a sequence complementary to SNA was added to the nanostructure, helicity was induced and a strong circular dichroism signal was observed. The SNA nanowire could amplify the helicity of chiral nucleic acids through nucleobase stacks. The SNA nanostructures have potential for use as platforms to detect chiral biomolecules under aqueous conditions because SNA can be readily functionalized and is water-soluble.

    DOI: 10.1039/d0sc05245k

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    PubMed

  12. Light-Regulated Liquid-Liquid Phase Separation for Spatiotemporal Protein Recruitment and Cell Aggregation 査読有り

    Ikeuchi N., Komachi T., Murayama K., Asanuma H., Maruyama A., Shimada N.

    ACS Applied Materials and Interfaces   13 巻 ( 4 ) 頁: 5652 - 5659   2021年2月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACS Applied Materials and Interfaces  

    We have previously shown that the upper critical solution temperature-type thermoresponsive ureido polymers such as polyallylurea and poly(2-ureidoethylmethacrylate) derivatives show liquid-liquid phase separation (LLPS), also known as simple coacervation, under physiological conditions below their phase-separation temperatures (Tp). The addition of the polymer-rich coacervate droplets that result from LLPS to a monolayer cell culture induced aggregation of cells into multicellular spheroids. In this study, we prepared a ureido copolymer, poly(vinylamine-co-vinylurea), with azobenzene substituents (Azo-PVU) and demonstrated light-guided assembly and disassembly of LLPS coacervates. Azo-PVUs with Tp values ranging from 10 to 52 °C were prepared by changing the azobenzene content. Ultraviolet light caused a decrease in the Tp of Azo-PVU because of trans-to-cis photoisomerization of the azobenzene and irradiation with visible light increased the Tp. Thus, LLPS of Azo-PVU was reversibly controlled. The coacervate droplets deposited on a dish surface were immediately dissolved by targeted UV irradiation (owing to a decrease in the Tp). Spatially controlled recruitment of proteins on the dish surface was achieved when protein solution was added to the light-patterned surface. Furthermore, the light-guided deposition of coacervates resulted in the spatiotemporal transformation of monolayer cells to aggregates. This light-controlled LLPS will allow the preparation of novel liquid-based materials for biomolecular and cellular engineering.

    DOI: 10.1021/acsami.0c22314

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    PubMed

  13. Quantitative Analyses of Förster Resonance Energy Transfer between Identical Pyrene Chromophores (Homo-FRET) In DNA Scaffolds. 査読有り

    Kashida H., Kawai H., Azuma H., Araki Y., Wada T., Asanuma H.

    ChemPhotoChem   5 巻 ( 2 ) 頁: 167 - 172   2021年2月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ChemPhotoChem  

    Förster resonance energy transfer between identical chromophores (homo-FRET) has been difficult to analyze since neither emission intensity nor lifetime changes with the occurrence of homo-FRET. Herein we used a DNA scaffold to analyze homo-FRET between pyrene moieties. The DNA scaffold was modified with two pyrenes and a quencher, anthraquinone. Homo-FRET was detected by monitoring quenching of pyrene emission and the decrease in the fluorescence lifetime of pyrene. Homo-FRET efficiencies could be calculated by excluding effects of hetero-FRET. The experimentally determined efficiencies showed an excellent agreement with Förster theory. These results will inform design of novel molecular probes and light-harvesting antennae.

    DOI: 10.1002/cptc.202000199

    Web of Science

    Scopus

  14. A Pyrene-Modified Serinol Nucleic Acid Nanostructure Converts the Chirality of Threoninol Nucleic Acids into Circularly Polarized Luminescence Signals 査読有り

    Kashida H., Nishikawa K., Ito Y., Murayama K., Hayashi I., Kakuta T., Ogoshi T., Asanuma H.

    Chemistry - A European Journal     2021年

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Chemistry - A European Journal  

    Herein is reported a circularly polarized luminescent (CPL) probe that can respond to the chirality of nucleic acids. An achiral nanostructure was prepared by the hybridization of symmetric serinol nucleic acid (SNA) containing pyrene-modified residues. When chiral oligomers that were complementary to the SNA were added, they induced helicity into the SNA nanowire. Efficient circular dichroism (CD) signal amplification was observed when pyrene was attached to uracil bases through a rigid alkynyl linker. Both CPL and CD signals were observed; they depended on the chirality of the added acyclic threoninol nucleic acid (aTNA) oligomer. This system can be used to convert the chirality of chiral biomolecules into chiroptical signals.

    DOI: 10.1002/chem.202102333

    Scopus

    PubMed

  15. RNA以前に非環状型XNAがPre-RNAとして存在したか? 招待有り 査読有り

    浅沼 浩之, 沖田 ひかり, 村山 恵司

    Viva Origino   49 巻 ( 3 ) 頁: 9   2021年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:生命の起原および進化学会  

    DOI: 10.50968/vivaorigino.49_9

    CiNii Research

  16. Intrastrand backbone-nucleobase interactions stabilize unwound right-handed helical structures of heteroduplexes of L-aTNA/RNA and SNA/RNA 査読有り

    Kamiya Y., Satoh T., Kodama A., Suzuki T., Murayama K., Kashida H., Uchiyama S., Kato K., Asanuma H.

    Communications Chemistry   3 巻 ( 1 ) 頁: 156   2020年12月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Communications Chemistry  

    Xeno nucleic acids, which are synthetic analogues of natural nucleic acids, have potential for use in nucleic acid drugs and as orthogonal genetic biopolymers and prebiotic precursors. Although few acyclic nucleic acids can stably bind to RNA and DNA, serinol nucleic acid (SNA) and L-threoninol nucleic acid (L-aTNA) stably bind to them. Here we disclose crystal structures of RNA hybridizing with SNA and with L-aTNA. The heteroduplexes show unwound right-handed helical structures. Unlike canonical A-type duplexes, the base pairs in the heteroduplexes align perpendicularly to the helical axes, and consequently helical pitches are large. The unwound helical structures originate from interactions between nucleobases and neighbouring backbones of L-aTNA and SNA through CH–O bonds. In addition, SNA and L-aTNA form a triplex structure via C:G*G parallel Hoogsteen interactions with RNA. The unique structural features of the RNA-recognizing mode of L-aTNA and SNA should prove useful in nanotechnology, biotechnology, and basic research into prebiotic chemistry.

    DOI: 10.1038/s42004-020-00400-2

    Web of Science

    Scopus

  17. Investigation of strand-selective interaction of SNA-modified siRNA with AGO2-MID 査読有り

    Kamiya Y., Takeyama Y., Mizuno T., Satoh F., Asanuma H.

    International Journal of Molecular Sciences   21 巻 ( 15 ) 頁: 1 - 13   2020年8月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:International Journal of Molecular Sciences  

    Small interfering RNA (siRNA) has been recognized as a powerful gene-silencing tool. For therapeutic application, chemical modification is often required to improve the properties of siRNA, including its nuclease resistance, activity, off-target effects, and tissue distribution. Careful siRNA guide strand selection in the RNA-induced silencing complex (RISC) is important to increase the RNA interference (RNAi) activity as well as to reduce off-target effects. The passenger strand-mediated off-target activity was previously reduced and on-target activity was enhanced by substitution with acyclic artificial nucleic acid, namely serinol nucleic acid (SNA). In the present study, the reduction of off-target activity caused by the passenger strand was investigated by modifying siRNAs with SNA. The interactions of SNA-substituted mononucleotides, dinucleotides, and (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO)-labeled double-stranded RNA (dsRNA) with the MID domain of the Argonaute 2 (AGO2) protein, which plays a pivotal role in strand selection by accommodation of the 5’-terminus of siRNA, were comprehensively analyzed. The obtained nuclear magnetic resonance (NMR) data revealed that AGO2-MID selectively bound to the guide strand of siRNA due to the inhibitory effect of the SNA backbone located at the 5’ end of the passenger strand.

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  18. Photo-regulated trajectories of gliding microtubules conjugated with DNA 査読有り

    Akter M., Keya J.J., Kabir A.M.R., Asanuma H., Murayama K., Sada K., Kakugo A.

    Chemical Communications   56 巻 ( 57 ) 頁: 7953 - 7956   2020年7月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Chemical Communications  

    We regulate the persistency in motion of kinesin-driven microtubules (MTs) simply using a photoresponsive DNA (pDNA) and ultraviolet (UV)-visible light. The path persistence length of MTs, which is a measure of the persistency in their motion, increases and decreases upon illuminating the MTs with UV and visible light respectively. Moreover, pDNA is found to work as a shield for MTs against damage under UV irradiation.

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  19. Efficient Light-Harvesting Antennae Resulting from the Dense Organization of Dyes into DNA Junctions through d-Threoninol 査読有り

    Kashida H., Azuma H., Maruyama R., Araki Y., Wada T., Asanuma H.

    Angewandte Chemie - International Edition   59 巻 ( 28 ) 頁: 11360 - 11363   2020年7月

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    担当区分:最終著者, 責任著者   記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Angewandte Chemie - International Edition  

    Herein we report the construction of efficient light-harvesting antennae by hybridization of DNA oligonucleotides containing high densities of fluorophores into DNA junctions through d-threoninol. Six pyrene donors could be incorporated into each arm without self-quenching. A perylene acceptor was located at the center of the junction. Antenna effects of a duplex and three- to eight-way junctions were systematically compared. Six- and eight-way junctions had the highest antenna effects, and their effective absorption coefficients were 8.5 times higher than that of perylene. Interestingly, even-numbered junctions had higher efficiencies than odd-numbered junctions. Nondenaturing gel analyses and fluorescence lifetime measurements demonstrated that the strong odd–even effects were derived from differences in the stability of junctions. The results presented will guide the design of efficient artificial photosynthetic systems.

    DOI: 10.1002/anie.202004221

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  20. A triplex-forming linear probe for sequence-specific detection of duplex DNA with high sensitivity and affinity 査読有り

    Chen Y., Murayama K., Kashida H., Kamiya Y., Asanuma H.

    Chemical Communications   56 巻 ( 40 ) 頁: 5358 - 5361   2020年5月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Chemical Communications  

    A triplex-forming oligonucleotide (TFO) linear probe containing perylene derivatives was synthesized. The TFO linear probe formed a remarkably stable triplex with a target DNA duplex, resulting in the light-up of fluorescence emission. The sensitivity was extremely high even at pH 7. Detection of PCR-amplified target DNA was demonstrated.

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  21. Incorporation of Pseudo-complementary Bases 2,6-Diaminopurine and 2-Thiouracil into Serinol Nucleic Acid (SNA) to Promote SNA/RNA Hybridization 査読有り

    Kamiya Y., Sato F., Murayama K., Kodama A., Uchiyama S., Asanuma H.

    Chemistry - An Asian Journal   15 巻 ( 8 ) 頁: 1266 - 1271   2020年4月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Chemistry - An Asian Journal  

    Serinol nucleic acid (SNA) is a promising candidate for nucleic acid-based molecular probes and drugs due to its high affinity for RNA. Our previous work revealed that incorporation of 2,6-diaminpurine (D), which can form three hydrogen bonds with uracil, into SNA increases the melting temperature of SNA-RNA duplexes. However, D incorporation into short self-complementary regions of SNA promoted self-dimerization and hindered hybridization with RNA. Here we synthesized a SNA monomer of 2-thiouracil (sU), which was expected to inhibit base pairing with D by steric hindrance between sulfur and the amino group. To prepare the SNA containing D and sU in high yield, we customized the protecting groups on D and sU monomers that can be readily deprotected under acidic conditions. Incorporation of D and sU into SNA facilitated stable duplex formation with target RNA by suppressing the self-hybridization of SNA and increasing the stability of the heteroduplex of SNA and its complementary RNA. Our results have important implications for the development of SNA-based probes and nucleic acid drugs.

    DOI: 10.1002/asia.201901728

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  22. A Quencher-Free Linear Probe from Serinol Nucleic Acid with a Fluorescent Uracil Analogue 査読有り

    Murayama K., Asanuma H.

    ChemBioChem   21 巻 ( 1-2 ) 頁: 120 - 128   2020年1月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ChemBioChem  

    With the goal of developing a quencher-free probe composed of an artificial nucleic acid, the fluorescent nucleobase analogue 5-(perylenylethynyl)uracil (PeU), which was incorporated into totally artificial serinol nucleic acid (SNA) as a substitute for thymine, has been synthesized. In the context of a 12-mer duplex with RNA, these fluorophores reduce duplex stability slightly compared with that of an SNA without PeU modification; thus suggesting that structural distortion is not induced by the modification. If two PeUs were incorporated at separate positions in an SNA, the fluorescent emission at λ≈490 nm was clearly enhanced upon hybridization with complementary RNA. A quencher-free SNA linear probe containing three PeUs, each separated by six nucleobases, has been designed. Detection of target RNA with high sensitivity and discrimination of a single-base mismatch has also been demonstrated.

    DOI: 10.1002/cbic.201900498

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  23. 8-Pyrenylvinyl Adenine Controls Reversible Duplex Formation between Serinol Nucleic Acid and RNA by [2 + 2] Photocycloaddition 査読有り

    Murayama K., Yamano Y., Asanuma H.

    Journal of the American Chemical Society   141 巻 ( 24 ) 頁: 9485 - 9489   2019年6月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Journal of the American Chemical Society  

    Photocontrol of duplex formation between the totally artificial serinol nucleic acid (SNA) and target RNA was made possible using a photoresponsive nucleobase 8-pyrenylvinyl adenine (PVA). PVA residues in SNA can be induced to undergo intrastrand [2 + 2] photocycloaddition by 455 nm light. Effective cycloreversion of the PVA photodimer results from irradiation with 340 nm light. These reactions occurred in high yield, rapidly, selectively, and reversibly. When the PVA-SNA/RNA duplex was irradiated with 455 nm light, almost complete dissociation of the duplex was attained, and 340 nm light restored duplex formation by cycloreversion. This is the first example of use of photocycloaddition and cycloreversion to photoregulate canonical duplex formation and dissociation reversibly at constant temperature. Thus, SNA bearing PVA residues have potential for use in photocontrollable biological tools targeting endogenous RNAs in cells as well as photodriven SNA machines.

    DOI: 10.1021/jacs.9b03267

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  24. cis-On/trans-Off of DNA Hybridization with Alkylthio-azobenzene on L-Threoninol Responding to Visible Light 招待有り 査読有り 国際共著

    Asanuma H., Ishikawa T., Yamano Y., Murayama K., Liang X.

    ChemPhotoChem   3 巻 ( 6 ) 頁: 418 - 424   2019年6月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ChemPhotoChem  

    An azobenzene modified with a highly branched secondary alkylthio group incorporated into DNA via an L-threoninol scaffold can serve as a photo-switch to reversibly control DNA hybridization. In the system reported here, irradiation with visible light at 400 nm causes photo-isomerization of trans-azobenzene to cis and 520 nm light causes the switch to the trans form. The DNA duplex is formed when the azobenzene is in the cis form (upon irradiation with 400 nm light) and is dissociated when the azobenzene is in the trans form (upon irradiation with 520 nm light). In our previously reported system, the switch required UV light. Here, visible light was used to induce both isomerizations, and a smooth trans-to-cis isomerization was observed below the melting temperature. This new photo-switch with visible light overcomes one of the limitations for the potential in vivo use of our method. Combining it with the previously reported photo-switch will allow the construction of elaborate DNA nanomachines.

    DOI: 10.1002/cptc.201900060

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  25. Orthogonally Photocontrolled Non-Autonomous DNA Walker 査読有り 国際共著

    Škugor M., Valero J., Murayama K., Centola M., Asanuma H., Famulok M.

    Angewandte Chemie - International Edition   58 巻 ( 21 ) 頁: 6948 - 6951   2019年5月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Angewandte Chemie - International Edition  

    There is considerable interest in developing progressively moving devices on the nanoscale, with the aim of using them as parts of programmable therapeutics, smart materials, and nanofactories. Present here is an entirely light-induced DNA walker based on orthogonal photocontrol. Implementing two azobenzene derivatives, S-DM-Azo and DM-Azo, enabled precise coordination of strand displacement reactions that powered a biped walker and guided it along a defined track in a non-autonomous way. This unprecedented type of molecular walker design offers high precision control over the movement in back-and-forth directions as desired, and is regulated solely by the sequence of the irradiation wavelengths. This concept may open new avenues for advancing non-autonomous progressive molecular motors, ultimately facilitating their application at the nanoscale.

    DOI: 10.1002/anie.201901272

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  26. Isothermal double-cycle catalytic system using DNAzyme and RNase H for the highly selective one-pot detection of oligonucleotides 査読有り 国際共著

    An R., Kawai H., Asanuma H., Komiyama M., Liang X.

    Analyst   144 巻 ( 8 ) 頁: 2773 - 2779   2019年4月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Analyst  

    With the use of a double-cycle system involving two catalytic reactions by RNase H and DNAzyme, the signal of oligoDNAs has been specifically amplified in an isothermal mode. The precursor of DNAzyme was introduced to the system as a ring-structured and inactivated form, which involves the 6-nt RNA portion being complementary to target oligoDNA. In the presence of target oligoDNA, the RNA portion forms a DNA/RNA hetero-duplex and is cut by RNase H. This scission converts the precursor to catalytically active DNAzyme, which in turn disconnects the molecular beacon to produce the amplified signal. Because the covalent bonds were disconnected to provide discrete structural changes in both cycles, high sensitivity and specificity are obtained, indicating the strong potential of this double catalytic cycle method for versatile applications.

    DOI: 10.1039/c8an02520g

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  27. N-Benzoyl-protected Peptide Nucleic Acid (PNA) Monomers Expand the Range of Nucleobases Available for PNA-DNA Chimera

    Inagaki M., Uematsu R., Mizutani T., Unabara D., Araki Y., Sakamoto S., Kashida H., Nishijima M., Asanuma H., Inoue Y., Wada T.

    Chemistry Letters   48 巻 ( 4 ) 頁: 341 - 344   2019年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 日本化学会  

    We synthesized N-benzoyl-protected peptide nucleic acid (PNA) monomers, which are robust under the conditions for deprotecting the 9-fluorenylmethoxycarbonyl (Fmoc) group by piperidine but are removable by aqueous ammonia and hence totally compatible with Fmoc-solid phase synthesis. This new invention expands the range of available nucleobase sequences, allowing us to use acid-sensitive PNA oligomers and purine nucleotides (both of which are difficult to use in the conventional methods) in the preparation of PNA-DNA chimeras to avoid the drawbacks of traditional PNAs.

    DOI: 10.1246/cl.181048

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  28. In-stem molecular beacon targeted to a 5 <sup>0</sup> -region of tRNA inclusive of the D arm that detects mature tRNA with high sensitivity 査読有り

    Miyoshi Y., Ohtsuki T., Kashida H., Asanuma H., Watanabe K.

    PLoS ONE   14 巻 ( 1 ) 頁: e0211505   2019年1月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PLoS ONE  

    Cellular functions are regulated by the up- and down-regulation and localization of RNA molecules. Therefore, many RNA detection methods have been developed to analyze RNA levels and localization. Molecular beacon (MB) is one of the major methods for quantitative RNA detection and analysis of RNA localization. Most oligonucleotide-based probes, including MB, are designed to target a long flexible region on the target RNA molecule, e.g., a single-stranded region. Recently, analyses of tRNA localization and levels became important, as it has been shown that environmental stresses and chemical reagents induce nuclear accumulation of tRNA and tRNA degradation in mammalian cells. However, tRNA is highly structured and does not harbor any long flexible regions. Hence, only a few methods are currently available for detecting tRNA. In the present study, we attempted to detect elongator tRNA Met (eMet) and initiator tRNA Met (iMet) by using an in-stem molecular beacon (ISMB), characterized by more effective quenching and significantly higher sensitivity than those of conventional MB. We found that ISMB1 targeted a 5 0 - region that includes the D arm of tRNA and that it detected eMet and iMet transcripts as well as mature eMet with high sensitivity. Moreover, the analysis revealed that the formation of the ISMB/tRNA transcript complex required more time than the formation of an ISMB/unstructured short RNA complex. These results suggest that ISMB-based tRNA detection can be a useful tool for various biological and medical studies.

    DOI: 10.1371/journal.pone.0211505

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  29. Photo-regulatable DNA isothermal amplification by template-mediated ligation 査読有り

    Cheng B., Kashida H., Shimada N., Maruyama A., Asanuma H.

    Chemical Communications   55 巻 ( 8 ) 頁: 1080 - 1083   2019年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Chemical Communications  

    By combining azobenzene-tethered oligonucleotides as modulators and poly(l-lysine)-graft-dextran (PLL-g-Dex), a chaperone polymer, to facilitate strand displacement, we successfully developed a photo-regulatable DNA isothermal amplification method. By alternating UV and visible irradiation, linear amplification was achieved. The method enables photo-regulatability and mismatch discrimination in linear amplification of the DNA target.

    DOI: 10.1039/c8cc09218d

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  30. Selective binding of nucleosides to gapped DNA duplex revealed by orientation and distance dependence of FRET 査読有り

    Kashida H., Kokubo Y., Makino K., Asanuma H.

    Organic and Biomolecular Chemistry   17 巻 ( 28 ) 頁: 6786 - 6789   2019年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Organic and Biomolecular Chemistry  

    Herein we used orientation and distance dependence of Förster resonance energy transfer (FRET) to analyze the binding of nucleosides to a gapped DNA duplex. Binding isotherms and information on the structures of the complexes were obtained by monitoring FRET between pyrene and perylene, which were introduced into the DNA through d-threoninol. FRET efficiency significantly changed upon formation of a duplex with a 1-nucleotide gap and a nucleoside. The FRET plot indicated that the complex has a double helical structure similar to a nicked duplex. Cooperative binding of two nucleosides to a duplex with a 2-nucleotide gap was also revealed using FRET. Various drug-nucleic acids interactions could be investigated using this sensitive and facile method.

    DOI: 10.1039/c9ob00946a

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  31. The DNA duplex as an aqueous one-Dimensional soft crystal scaffold for photochemistry 招待有り

    Asanuma H., Murayama K., Kamiya Y., Kashida H.

    Bulletin of the Chemical Society of Japan   91 巻 ( 12 ) 頁: 1739 - 1748   2018年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 日本化学会  

    In this account, we demonstrate that DNA duplex is an ideal scaffold for photochemistry, particularly for comparison of photochemical theory with experiments. The well-defined structure of a DNA duplex can be regarded as an aqueous one-dimensional soft crystal composed of a chromophore-like base-pair assembly. When any base pair in the duplex is replaced with a chromophore, orientation, distance, and association number of chromophores can be precisely controlled. We have developed a new methodology for introduction of chromophores into DNA duplexes using D-threoninol. By using the DNA duplex as a scaffold, experiments on exciton interactions of chromophore assemblies can be compared with molecular exciton theory. A fluorescent resonance energy transfer (FRET) system was also constructed by introducing donor pyrene and acceptor perylene into the DNA duplex using D-threoninol monomers. Using this system, we demonstrated orientation-dependent FRET. We found that theories on both exciton interaction and FRET qualitatively coincide with experimental data and revealed the limitation of the point-dipole approximation. We also evaluated the intrinsic quantum yield of photodimerization of stilbene derivatives by suppressing a side reaction. We propose that there is a correlation of quantum yield of photodimerization with the energy gap of HOMO or LUMO, a hypothesis that deserves theoretical investigation.

    DOI: 10.1246/bcsj.20180278

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  32. Quantitative evaluation of energy migration between identical chromophores enabled by breaking symmetry 査読有り

    Kashida H., Kawai H., Maruyama R., Kokubo Y., Araki Y., Wada T., Asanuma H.

    Communications Chemistry   1 巻 ( 1 )   2018年12月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Communications Chemistry  

    Energy migration between the identical chromophores is a necessary process in both natural and artificial photosynthesis. The distance and orientation dependence of energy migration have not been experimentally investigated in detail. Here we propose a method to investigate energy migration. Two fluorophores are introduced into one strand of a DNA duplex with a quencher placed opposite one of fluorophores. This design enables asymmetrization of identical fluorophores and allows one fluorophore to behave as an acceptor. The emission intensities and lifetimes decrease depending on the efficiency of energy migration. Distance and orientation dependence are successfully quantified, and the excitation energy migration efficiencies measured are in excellent agreement with those calculated based on Förster theory. We also demonstrate that multi-step energy migration among four fluorophores can be estimated from the theory. These results may provide a basis for design and preparation of efficient light-harvesting photonic devices and chemical probes.

    DOI: 10.1038/s42004-018-0093-0

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  33. Bifacial Nucleobases for Hexaplex Formation in Aqueous Solution 査読有り

    Kashida H., Hattori Y., Tazoe K., Inoue T., Nishikawa K., Ishii K., Uchiyama S., Yamashita H., Abe M., Kamiya Y., Asanuma H.

    Journal of the American Chemical Society   140 巻 ( 27 ) 頁: 8456 - 8462   2018年7月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Journal of the American Chemical Society  

    Although DNA can form triplex and quadruplex structures through hydrogen bonds, design and preparation of structures with more than five strands is difficult even when artificial nucleic acids are used. Herein we report a hexaplex formed by oligomers of artificial nucleic acids bearing bifacial molecules on d-threoninol. Aminopyrimidine and cyanuric acid derivatives were selected as bases because they have complementary hydrogen bonding patterns. The complex formed by aminopyrimidine and cyanuric acid decamers melted with large hysteresis. Hexaplex formation was indicated by gel electrophoresis, size exclusion chromatography and atomic force microscopy imaging, and proven directly through native mass spectrometry. CD measurements and molecular dynamics simulations indicated that the hexaplex adopts a helical structure. The hexaplex formation was highly dependent on pH and the presence of divalent cations. The hexaplex was stable in aqueous solution, and its unique structure and properties may lead to novel nanostructures, molecular assemblies, metal sensors, and ion channels.

    DOI: 10.1021/jacs.8b02807

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  34. DNA-assisted swarm control in a biomolecular motor system 査読有り

    Keya Jakia Jannat, Suzuki Ryuhei, Kabir Arif Md. Rashedul, Inoue Daisuke, Asanuma Hiroyuki, Sada Kazuki, Hess Henry, Kuzuya Akinori, Kakugo Akira

    NATURE COMMUNICATIONS   9 巻   頁: 453   2018年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1038/s41467-017-02778-5

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  35. Does Treatment of Impaired Glucose Tolerance Improve Cardiovascular Outcomes in Patients with Previous Myocardial Infarction?

    Asakura M., Kim J., Asanuma H., Hamasaki T., Tsukahara K., Higashino Y., Ishikawa T., Nakama Y., Koba S., Maruyama Y., Tsujimoto M., Himeno H., Ohkusa T., Fujino S., Shimizu M., Endo T., Yoda S., Muroya T., Murohara T., Ohte N., Suzuki H., Kohno T., Fukui K., Shiono T., Takase H., Uzui H., Nagai Y., Hashimoto Y., Ikeda S., Mizuno S., Tamita K., Fujita M., Satake K., Kinoshita Y., Nunohiro T., Sakagami S., Higaki J., Morii I., Sawada R., Hiasa Y., Shigemasa T., Nakahama M., Sata M., Doi O., Ueda T., Yamada T., Yamanouchi T., Yamaguchi H., Morita Y., Hayashi H., Kitakaze M.

    Cardiovascular Drugs and Therapy   31 巻 ( 4 ) 頁: 401 - 411   2017年8月

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    記述言語:日本語   出版者・発行元:Cardiovascular Drugs and Therapy  

    Purpose: We evaluated the effects of an alpha-glucosidase inhibitor, voglibose, on cardiovascular events in patients with a previous myocardial infarction (MI) and impaired glucose tolerance (IGT). Methods: This prospective, randomized, open, blinded-endpoint study was conducted in 112 hospitals and clinics in Japan in 3000 subjects with both previous MI and IGT receiving voglibose (0.6 mg/day, n = 424) or no drugs (n = 435) for 2 years. The Data and Safety Monitoring Board (DSMB) recommended discontinuation of the study in June 2012 after an interim analysis when the outcomes of 859 subjects were obtained. The primary endpoint was cardiovascular events including cardiovascular death, nonfatal MI, nonfatal unstable angina, nonfatal stroke, and percutaneous coronary intervention/coronary artery bypass graft. Secondary endpoints included individual components of the primary endpoint in addition to all-cause mortality and hospitalization due to heart failure. Results: The age, ratio of males, and HbA1C were 65 vs. 65 years, 86 vs. 87%, and 5.6 vs. 5.5% in the groups with and without voglibose, respectively. Voglibose improved IGT; however, Kaplan–Meier analysis showed no significant between-group difference with respect to cardiovascular events [12.5% with voglibose vs. 10.1% without voglibose for the primary endpoint (95% confidence interval, 0.82–1.86)]; there were no significant differences in secondary endpoints. Conclusion: Although voglibose effectively treated IGT, no additional benefits for cardiovascular events in patients with previous MI and IGT were observed. Voglibose may not be a contributing therapy to the secondary prevention in patients with MI and IGT. Trial Registration: Clinicaltrials.gov number: NCT00212017.

    DOI: 10.1007/s10557-017-6740-3

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  36. Antisense oligonucleotide modified with serinol nucleic acid (SNA) induces exon skipping in: Mdx myotubes

    Le B., Murayama K., Shabanpoor F., Asanuma H., Veedu R.

    RSC Advances   7 巻 ( 54 ) 頁: 34049 - 34052   2017年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:RSC Advances  

    Serinol nucleic acid (SNA) is a novel nucleic acid analogue that can form highly stable heteroduplexes with complementary DNA and RNA sequences. Structurally, SNA is a close mimic to peptide nucleic acid (PNA) which is widely used in diagnostic and therapeutic applications. SNA chemistry is relatively new, and so far the scope of SNA has only been explored in improving the efficacy of small interfering RNA and for developing a highly sensitive molecular beacon for diagnostic applications. In this study, we investigated the potential of SNA-modified antisense oligonucleotide (AO) in parallel to PNA-oligo for splice-modulation in an in vitro cellular model of Duchenne muscular dystrophy (DMD). We synthesized a 20-mer SNA and PNA antisense oligonucleotide (AO) designed to induce exon-23 skipping in the mouse dystrophin gene transcript. Our results demonstrated that the SNA AO induced exon-23 skipping at all tested concentrations, whereas the corresponding PNA AO failed to induce any exon-23 skipping upon 24 hours of transfection using Lipofectin transfection reagent. Our results further expands the potential of SNA oligonucleotides in therapeutic applications.

    DOI: 10.1039/c7ra06091b

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  37. Development of Pseudo Base-Pairs on D-Threoninol which Exhibit Various Functions

    Kashida Hiromu, Asanuma Hiroyuki

    Bulletin of the Chemical Society of Japan   90 巻 ( 5 ) 頁: 475 - 484   2017年

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    記述言語:英語   出版者・発行元:公益社団法人 日本化学会  

    <p>The authors have developed various kinds of pseudo base pairs using a <span style="font-variant: small-caps;">d</span>-threoninol scaffold. Although the chemical structures of the pseudo base pairs are much different from natural nucleobases, they can mimic supramolecular properties of natural base pairs. Moreover, modified DNA can possess various functions that cannot be achieved by natural nucleic acids, such as fluorescent switchability, photocrosslinking, insulating and emission color change. These pseudo base pairs can be used to prepare various functional nanomaterials. In the present account, we summarize our recent work on pseudo base pairs, focusing on molecular designs and functions.</p>

    DOI: 10.1246/bcsj.20160371

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  38. Visible-Light-Triggered Cross-Linking of DNA Duplexes by Reversible [2+2] Photocycloaddition of Styrylpyrene

    Doi T., Kawai H., Murayama K., Kashida H., Asanuma H.

    Chemistry - A European Journal   22 巻 ( 30 ) 頁: 10533 - 10538   2016年7月

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    記述言語:日本語   出版者・発行元:Chemistry - A European Journal  

    Reversible photo-cross-linking of a DNA duplex through the [2+2] photocycloaddition of styrylpyrene is reported. Styrylpyrene moieties on d-threoninol linkers were introduced into complementary positions on DNA strands. Irradiation of the styrylpyrene pair in the duplex with visible light at λ=455 nm induced a [2+2] photocycloaddition between styrylpyrenes that cross-linked the two strands of the duplex. Two diastereomers were formed after [2+2] photocycloaddition as a result of rotation of the styrylpyrene residues. Also, the cycloreversion reaction was induced by UV light at λ=340 nm, which reversibly yielded the uncross-linked strands.

    DOI: 10.1002/chem.201602006

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  39. Faint electric treatment-induced rapid and efficient delivery of extraneous hydrophilic molecules into the cytoplasm

    Hasan M., Nishimoto A., Ohgita T., Hama S., Kashida H., Asanuma H., Kogure K.

    Journal of Controlled Release   228 巻   頁: 20 - 25   2016年4月

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    記述言語:日本語   出版者・発行元:Journal of Controlled Release  

    Effective delivery of extraneous molecules into the cytoplasm of the target cells is important for several drug therapies. Previously, we showed effective in vivo transdermal delivery of naked siRNA into skin cells induced by faint electric treatment (ET) iontophoresis, and significant suppression of target mRNA levels (Kigasawa et al., Int. J. Pharm., 2010). This result indicates that electricity promoted the delivery of siRNA into cytoplasm. In the present study, we analyzed the intracellular delivery of naked anti-luciferase siRNA by faint ET, and found that the luciferase activity of cells expressing luciferase was reduced by in vitro ET like in vivo iontophoresis. Cellular uptake of fluorescent-label siRNA was increased by ET, while low temperature exposure, macropinocytosis inhibitor amiloride and caveolae-mediated endocytosis inhibitor filipin significantly prevented siRNA uptake. These results indicate that the cellular uptake mechanism involved endocytosis. In addition, voltage sensitive fluorescent dye DiBAC4 (3) penetration was increased by ET, and the transient receptor potential channel inhibitor SKF96365 reduced siRNA uptake, suggesting that faint ET reduced membrane potentials by changing intracellular ion levels. Moreover, to analyze cytoplasmic delivery, we used in-stem molecular beacon (ISMB), which fluoresces upon binding to target mRNA in the cytoplasm. Surprisingly, cytoplasmic ISMB fluorescence appeared rapidly and homogeneously after ET, indicating that cytoplasmic delivery is markedly enhanced by ET. In conclusion, we demonstrated for the first time that faint ET can enhance cellular uptake and cytoplasmic delivery of extraneous molecules.

    DOI: 10.1016/j.jconrel.2016.02.048

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  40. 天然に存在しない非環状構造の核酸アナログ-人工の非環状骨格で神の設計した環状骨格に挑む

    浅沼 浩之

    化学   71 巻   頁: 2016   2016年

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  41. Terminus-free siRNA prepared by photo-crosslinking activated via slicing by Ago2

    Kamiya Y., Iishiba K., Doi T., Tsuda K., Kashida H., Asanuma H.

    Biomaterials Science   3 巻 ( 12 ) 頁: 1534 - 1538   2015年12月

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    記述言語:日本語   出版者・発行元:Biomaterials Science  

    We report the development of photo-crosslinked siRNA strands modified at each terminus with p-cyanostilbene. The siRNA was nuclease resistant and retained RNAi activity. We further studied the activation mechanism of the covalently-crosslinked siRNA. Interestingly Dicer, which is known to generate siRNA with overhanging 3′ ends from the precursor siRNA, did not cleave the crosslinked siRNA at all. Our results suggest that the activation of the crosslinked siRNAs required cleavage by Argonaute2.

    DOI: 10.1039/c5bm00231a

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  42. Pre-organized Guide RNA in the Cas9 Complex Is Ready for the Selection of Target Double-Stranded DNA

    Kamiya Y., Asanuma H.

    ChemBioChem   16 巻 ( 16 ) 頁: 2273 - 2275   2015年11月

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    記述言語:日本語   出版者・発行元:ChemBioChem  

    The 3 D structure of Cas9 bound with sgRNA was solved by X-ray crystal-structure analysis. The conformation change in SpyCas9 upon binding to sgRNA changes Cas9 to target-DNA-recognition mode in which seed segments in sgRNA are pre-ordered in an A-type conformation.

    DOI: 10.1002/cbic.201500424

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  43. Detection of DNA target with highly enhanced specificity by self-circularization rolling circle amplification

    Wang X., Wang X., Dong P., Suzuki M., Asanuma H., Liang X.

    Lecture Notes in Electrical Engineering   251 LNEE 巻 ( VOL. 3 ) 頁: 1449 - 1458   2014年2月

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    記述言語:日本語   出版者・発行元:Lecture Notes in Electrical Engineering  

    DNA detection is widely used for gene analysis. However, false-positive results are difficult to be avoided due to the non-specific amplification. Here, we described a novel RCA approach to improve the specificity. With the help of TspR I (endonuclease), DNA was cleaved into smaller duplex fragments, including the target DNA fragment, with 9 nt sticky ends. A duplex adaptor with two sticky ends which were complementary to that of the target fragment was designed. After hybridization of the adaptor with the target fragments, T4 DNA ligase was used to ligate them to form a circular ds DNA with a gap. Then RCA was carried out from the free 3′-end of the open strand by Phi29 DNA polymerase with two primers which were complementary to the target sequence. Different from the traditional padlock-RCA, SC-RCA showed excellent specificity by amplifying the specific DNA target other than the added probe. © Springer-Verlag Berlin Heidelberg 2014.

    DOI: 10.1007/978-3-642-37925-3_154

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  44. SNA: A nucleic acid analogue that can control the helical structure by sequence design

    Asanuma H., Murayama K., Kashida H.

    Kobunshi   62 巻 ( 9 ) 頁: 513 - 514   2013年9月

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    記述言語:日本語   出版者・発行元:Kobunshi  

    We synthesized a unique nucleic acid analogue, Serinol Nucleic Acid (SNA), from a serinol scaffold tethering natural nucleobases through an amide bond. The SNA oligomer that is fully synthesized from four chiral SNA monomers changes its chirality by the sequence design, not by the inherent chirality of monomers: The SNA oligomer with an asymmetric sequence is chiral whereas the symmetric one is achiral. The chirality of the SNA oligomer can be inverted by reversing its sequence, i.e., two enantiomers of SNA oligomers can be synthesized from four monomers with the identical chirality. Interestingly, SNA formed a remarkably stable duplex with a complementary SNA in an antiparallel manner with its helicity depending on the sequence, which is much more stable than the corresponding DNA/DNA or RNA/RNA duplex. More interestingly, SNA also formed a stable duplex both with DNA and RNA, indicating high potential for antisense agents. These unique properties of SNA might provide an insight for why D-ribose was selected as a scaffold for natural nucleic acids. © 2013 The Society of Polymer Science, Japan.

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  45. A “sugar-deficient” G-quadruplex: Incorporation of aTNA in G4 structures

    Zhou J., Murayama K., Amrane S., Rosu F., Kashida H., Bourdoncle A., Asanuma H., Mergny J.L.

    Chemical Science   4 巻 ( 9 ) 頁: 3693 - 3698   2013年7月

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    記述言語:日本語   出版者・発行元:Chemical Science  

    The effects of modification of the phosphodiester backbone or the guanine bases on G-quadruplex formation have been widely investigated. Only a few studies have investigated the effects of deoxyribose or ‘sugar’ modifications on G-quadruplex structure. Here, we evaluated the structural, thermodynamic, and kinetic properties of the parallel quadruplexes formed by the sequence d(TGGGGT) in which each guanine base was substituted, one at a time, with acyclic threoninol nucleic acid (aTNA). We found that all sequences were able to form G-quadruplexes; however, the presence of aTNA resulted in the formation of a mixture of quadruplex structures in some cases. Furthermore, the presence of a single substitution at any position resulted in destabilization of the G-quadruplex relative to that formed by the unmodified sequence. The introduction of the aTNA in terminal quartets was the most detrimental to stability. In addition, kinetic experiments showed that, compared to its unmodified counterpart sequence d(TGGGGT), the substitution of a normal guanine nucleotide by aTNA decelerated quadruplex formation except when the aTNA was at the 5′ most guanine of the sequence. In summary, our studies indicate that the deoxyribose sugar affects the properties of G-quadruplex structures. © 2013 The Royal Society of Chemistry.

    DOI: 10.1039/c3sc50474c

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  46. P-stilbazole moieties as artificial base pairs for photo-cross-linking of DNA duplex

    Kashida H., Doi T., Sakakibara T., Hayashi T., Asanuma H.

    Journal of the American Chemical Society   135 巻 ( 21 ) 頁: 7960 - 7966   2013年5月

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    記述言語:日本語   出版者・発行元:Journal of the American Chemical Society  

    In this study, we report a photo-cross-linking reaction between p-stilbazole moieties. p-Stilbazoles were introduced into base-paring positions of complementary DNA strands. The [2 + 2] photocycloaddition reaction occurred rapidly upon light irradiation at 340 nm. Consequently, duplex was cross-linked and highly stabilized after 3 min irradiation. The CD spectrum of the cross-linked duplex indicated that the B-form double-helical structure was not severely distorted. NMR analysis revealed only one conformation of the duplex prior to UV irradiation, whereas two diastereomers were detected after the photo-cross-linking reaction. Before UV irradiation, p-stilbazole can adopt two different stacking modes because of rotation around the single bond between the phenyl and vinyl groups; these conformations cannot be discriminated on the NMR time scale due to rapid interconversion. However, photo-cross-linking fixed the conformation and enabled discrimination both by NMR and HPLC. The artificial base pair of p-methylstilbazolium showed almost the same reactivity as p-stilbazole, indicating that positive charge does not affect the reactivity. When a natural nucleobase was present in the complementary strand opposite p-stilbazole, the duplex was significantly destabilized relative to the duplex with paired p-stilbazole moieties and no photoreaction occurred between p-stilbazole and the nucleobase. The p-stilbazole pair has potential as a "third base pair" for nanomaterials due to its high stability and superb orthogonality. © 2013 American Chemical Society.

    DOI: 10.1021/ja401835j

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  47. Quencher-free linear probe with multiple fluorophores on an acyclic scaffold

    Asanuma H., Akahane M., Kondo N., Osawa T., Kato T., Kashida H.

    Chemical Science   3 巻 ( 11 ) 頁: 3165 - 3169   2012年11月

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    記述言語:日本語   出版者・発行元:Chemical Science  

    We have developed a new quencher-free stemless linear probe involving multiple perylenes incorporated through d-threoninol; each perylene is separated by intervening natural nucleotides. Without a substrate, the flexible linear probe does not emit fluorescence due to the self-quenching of the weakly interacting fluorophores. Upon hybridization with the target, intercalation of each dye between the base pairs results in emission of strong fluorescence. The maximum signal-background ratio attained was 180, and the response rate was significantly faster than that of a classic hairpin-forming molecular beacon. © 2012 The Royal Society of Chemistry.

    DOI: 10.1039/c2sc20732j

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  48. Reversed assembly of dyes in an RNA duplex compared with those in DNA

    Fujii T., Urushihara M., Kashida H., Ito H., Liang X., Yagi-Utsumi M., Kato K., Asanuma H.

    Chemistry - A European Journal   18 巻 ( 42 ) 頁: 13304 - 13313   2012年10月

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    記述言語:日本語   出版者・発行元:Chemistry - A European Journal  

    We prepared reversed dye clusters by hybridizing two RNA oligomers, each of which tethered dyes (Methyl Red, 4'-methylthioazobenzene, and thiazole orange) on D-threoninols (threoninol nucleotides) at the center of their strands. NMR spectroscopic analyses revealed that two dyes from each strand were axially stacked in an antiparallel manner to each other in the duplex, and were located adjacent to the 3'-side of a natural nucleobase. Interestingly, this positional relationship of the dyes was completely the opposite of that assembled in DNA that we reported previously: dyes in DNA were located adjacent to the 5'-side of a natural nucleobase. This observation was also consistent with the circular dichroism of dimerized dyes in which the Cotton effect of the dyes (i.e., the winding properties of two dyes) was inverted in RNA relative to that in DNA. Further spectroscopic analyses revealed that clustering of the dyes on RNA duplexes induced distinct hypsochromicity and narrowing of the band, thus demonstrating that the dyes were axially stacked (i.e., H-aggregates) even on an A-type helix. On the basis of these results, we also prepared heterodimers of a fluorophore (thiazole orange) and quencher (Methyl Red) in an RNA duplex. Fluorescence from thiazole orange was found to be strongly quenched by Methyl Red due to the excitonic interaction, so that the ratio of fluorescent intensities of the RNA-thiazole orange conjugate with and without its complementary strand carrying a quencher became as high as 27. We believe that these RNA-dye conjugates are potentially useful probes for real-time monitoring of RNA interference (RNAi) mechanisms. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

    DOI: 10.1002/chem.201201956

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  49. Model of elongation of short DNA sequence by thermophilic DNA polymerase under isothermal conditions

    Kato T., Liang X., Asanuma H.

    Biochemistry   51 巻 ( 40 ) 頁: 7846 - 7853   2012年10月

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    記述言語:日本語   出版者・発行元:Biochemistry  

    Short DNA sequences, especially those that are repetitive or palindromic, can be used as the seeds for synthesis of long DNA by some DNA polymerases in an unusual manner. Although several elongation mechanisms have been proposed, there is no well-established model that explains highly efficient elongation under isothermal conditions. In the present study, we analyzed the elongation of nonrepetitive sequences with distinct hairpins at each end. These DNAs were elongated efficiently under isothermal conditions by thermophilic Vent (exo -) DNA polymerase, and the products were longer than 10 kb within 10 min of the reaction. A 20-nucleotide DNA with only one hairpin was also elongated. Sequence analysis revealed that the long products are mainly tandem repeats of the short seed sequences. The thermal melting temperatures of the products were much higher than the reaction temperature, indicating that most DNAs form duplexes during the reaction. Accordingly, a terminal hairpin formation and self-priming extension model was proposed in detail, and the efficient elongation was explained. Formation of the hairpin at the 5′ end plays an important role during the elongation. © 2012 American Chemical Society.

    DOI: 10.1021/bi3010413

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  50. A photon-fueled DNA nanodevice that contains two different photoswitches

    Nishioka H., Liang X., Kato T., Asanuma H.

    Angewandte Chemie - International Edition   51 巻 ( 5 ) 頁: 1165 - 1168   2012年1月

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    記述言語:日本語   出版者・発行元:Angewandte Chemie - International Edition  

    DNA seesaw: Photoswitchable azobenzenecarboxylic acid 1 reversibly photoisomerizes between the trans form and the thermally stable cis form upon irradiation with visible light. A photon-fueled DNA nanodevice that moves like a seesaw in response to irradiation with different wavelengths of light was made by modifying DNA oligonucleotides with a combination of 1 and a conventional azobenzene (see picture). Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

    DOI: 10.1002/anie.201106093

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  51. A polycation-chaperoned in-stem molecular beacon system

    Asanuma H., Osawa T., Kashida H., Fujii T., Liang X., Niwa K., Yoshida Y., Shimada N., Maruyama A.

    Chemical Communications   48 巻 ( 12 ) 頁: 1760 - 1762   2012年1月

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    記述言語:日本語   出版者・発行元:Chemical Communications  

    In the presence of poly(l-lysine)-graft-dextran, an in-stem molecular beacon involving three perylene–anthraquinone pairs in the stem region had a signal/background ratio of as high as 570. Response speed was also remarkable; equilibrium was attained within 5 minutes after addition of substrate DNA at 20 °C. © 2012 The Royal Society of Chemistry.

    DOI: 10.1039/c2cc16812j

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  52. Nick sealing by T4 DNA ligase on a modified DNA template: Tethering a functional molecule on D -threoninol

    Liang X., Fujioka K., Asanuma H.

    Chemistry - A European Journal   17 巻 ( 37 ) 頁: 10388 - 10396   2011年9月

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    記述言語:日本語   出版者・発行元:Chemistry - A European Journal  

    Efficient DNA nick sealing catalyzed by T4 DNA ligase was carried out on a modified DNA template in which an intercalator such as azobenzene had been introduced. The intercalator was attached to a D-threoninol linker inserted into the DNA backbone. Although the structure of the template at the point of ligation was completely different from that of native DNA, two ODNs could be connected with yields higher than 90 % in most cases. A systematic study of sequence dependence demonstrated that the ligation efficiency varied greatly with the base pairs adjacent to the azobenzene moiety. Interestingly, when the introduced azobenzene was photoisomerized to the cis form on subjection to UV light (320-380 nm), the rates of ligation were greatly accelerated for all sequences investigated. These unexpected ligations might provide a new approach for the introduction of functional molecules into long DNA strands in cases in which direct PCR cannot be used because of blockage of DNA synthesis by the introduced functional molecule. The biological significance of this unexpected enzymatic action is also discussed on the basis of kinetic analysis,. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

    DOI: 10.1002/chem.201100215

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  53. A cationic dye triplet as a unique "glue" that can connect fully matched termini of DNA duplexes

    Kashida H., Hayashi T., Fujii T., Asanuma H.

    Chemistry - A European Journal   17 巻 ( 9 ) 頁: 2614 - 2622   2011年2月

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    記述言語:日本語   出版者・発行元:Chemistry - A European Journal  

    In this study, we propose that three consecutive cationic p-methylstilbazoles tethered on D-threoninols (Z residues) at 5′ termini act as a unique "glue" connecting DNA duplexes by their interstrand cluster formation. Interstrand clustering of p-methylstilbazoles (ZZZ triplets) induces narrowing and hypsochromic shift of bands at 350nm, which can be assigned to the absorption of p-methylstilbazole. However, single-stranded DNA conjugates involving a ZZZ triplet at the 5′ terminus of 8-mer native nucleotides is found not to induce such large spectral changes, which implies that the intrinsic self-assembling property of ZZZ triplets is weak. Interestingly, when this conjugate is hybridized with a complementary 8-mer native oligonucleotide, a remarkable spectral change is observed, indicating the dimerization of a duplex through the interstrand clustering of ZZZ triplets. Dimerization of the duplex is also evidenced by cold-spray ionization mass spectrometry. This interstrand clustering is observed only when a ZZZ triplet is tethered to a 5′ rather than 3′ terminus. Furthermore, the stability of the interstrand cluster increases by increasing the number of nucleobases of the DNA portion, and when mismatched base pairs are incorporated or when a base next to the Z residue is deleted, the stability substantially drops. When we apply the ZZZ triplet to the formation of a nanowire using two complementary DNA conjugates, each of which has a ZZZ triplet at the 5′ termini as overhang, we demonstrate the successful formation of a nanowire by native PAGE analysis. Since native sticky ends that have three nucleotides do not serve as "glue", ZZZ triplets with their unique glue-like properties are prime candidates for constructing DNA-based nanoarchitectures. Stick together! A unique "glue" of cationic p-methylstilbazoles tethered on D-threoninols connects DNA duplexes by intermolecular clustering. When the "glue" is attached to both termini of a DNA duplex, a nanowire is formed (see graphic). Thus, the "glue" can be utilized for constructing DNA-based nanoarchitectures. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

    DOI: 10.1002/chem.201003059

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  54. Design of a functional nanomaterial with recognition ability for constructing light-driven nanodevices

    Liang X., Mochizuki T., Fujii T., Kashida H., Asanuma H.

    Lecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics)   6518 LNCS 巻   頁: 112 - 122   2011年2月

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    記述言語:日本語   出版者・発行元:Lecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics)  

    An artificial macromolecule (foldamer) was designed as a novel nanomaterial with the backbone of phosphodiester and the side chain of functional molecules and nucleobases. The functional molecules tethered on D-threoninol and the nucleosides on D-ribose can be lined up with any sequence and ratio by using standard phosphoramidite chemistry. The nucleobases that form Watson-Crick base pairs provide the sequence recognition which is required for constructing complicate nanostructures. The multiple functional molecules give applicable and advanced functions such as photoresponsiveness when azobenzenes were used. Unexpectedly, a stable double helix was formed even in the case that the ratio of azobenzene molecules and base pairs was as high as 2:1. More interestingly, this artificial duplex showed high sequence specificity: the stability decreased greatly when a mismatched base pair was present. Furthermore, the formation and dissociation of the constructed artificial duplex were reversibly and completely modulated with light irradiation. By using this new nanomaterial, a variety of functional nanostructures and nanodevices are promising to be designed. © 2011 Springer-Verlag.

    DOI: 10.1007/978-3-642-18305-8_11

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  55. A light-driven DNA nanomachine for the efficient photoswitching of RNA digestion

    Zhou M., Liang X., Mochizuki T., Asanuma H.

    Angewandte Chemie - International Edition   49 巻 ( 12 ) 頁: 2167 - 2170   2010年3月

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    記述言語:日本語   出版者・発行元:Angewandte Chemie - International Edition  

    "Chemical Equation Presented" Photons as fuel: A photoresponsive DNA enzyme was constructed that can work at the single-molecule level. Complete ONOFF photoswitching of RNA digestion was realized by photoregulating the topological structure of a DNAzyme/RNA complex. The key components of the photoswitch are azobenzene units. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.

    DOI: 10.1002/anie.200907082

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  56. Photoregulation of DNA transcription by using photoresponsive T7 promoters and clarification of its mechanism

    Liang X., Wakuda R., Fujioka K., Asanuma H.

    FEBS Journal   277 巻 ( 6 ) 頁: 1551 - 1561   2010年3月

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    記述言語:日本語   出版者・発行元:FEBS Journal  

    With the use of photoresponsive T7 promoters tethering two 2′-methylazobenzenes or 2′,6′-dimethylazobenzenes, highly efficient photoregulation of DNA transcription was obtained. After UV-A light irradiation (320-400 nm), the rate of transcription with T7 RNA polymerase and a photoresponsive promoter involving two 2′,6′-dimethylazobenzenes was 10-fold faster than that after visible light irradiation (400-600 nm). By attaching a nonmodified azobenzene and 2′,6′-dimethylazobenzene at the two positions, respectively, and by utilizing the different cis→trans thermal stability between cis-nonmodified azobenzene and cis-2′,6′- dimethylazobenzene, four species of T7 promoter (cis-cis, trans-cis, cis-trans, and trans-trans) were obtained. The four species showed transcriptional activity in the order of cis-cis > cis-trans > trans-cis > trans-trans. Kinetic analysis revealed that the Km for the cis-cis promoter (both of the introduced azobenzene derivatives were in the cis form) and T7 RNA polymerase was 68 times lower than that for the trans-trans form, indicating that high photoregulatory efficiency was mainly due to a remarkable difference in affinity for RNA polymerase. The present approach is promising for the creation of biological tools for artificially controlling gene expression, and as a photocontrolled system for supplying RNA fuel for RNA-powered molecular nanomachines. © 2010 FEBS.

    DOI: 10.1111/j.1742-4658.2010.07583.x

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  57. Molecular imprinting of cyclodextrin to physiologically active oligopeptides in water

    Song S., Shirasaka K., Hirokawa Y., Asanuma H., Wada T., Sumaoka J., Komiyama M.

    Supramolecular Chemistry   22 巻 ( 3 ) 頁: 149 - 155   2010年3月

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    記述言語:日本語   出版者・発行元:Supramolecular Chemistry  

    β-Cyclodextrin (β-CyD)-based polymeric receptors for γ-endorphin (γ-endor, an opioid heptadecapeptide) were prepared using the molecular imprinting method. When mono-3-(N-acrylamido)-3-deoxy-β-CyD bearing a vinyl group in the secondary hydroxyl side of the cavity of β-CyD was polymerised in water in the presence of γ-endor, the binding activity of the β-CyD polymer to this peptide in water was enormously promoted by the imprinting. By contrast, the bindings towards methionine-enkephalin (N-terminal pentapeptide of γ-endor) and its homologue leucine-enkephalin were suppressed. Thus, the binding of γ-endor by the imprinted polymer was highly selective. The imprinting towards γ-endor was also successful with the use of the β-CyD monomer bearing a vinyl group in the primary hydroxyl side of the cavity, although the recognition was less strict. Various factors affecting the imprinting efficiency (kinds of β-CyD vinyl monomer and template, as well as the pH of imprinting mixture) are discussed. © 2010 Taylor & Francis.

    DOI: 10.1080/10610270902980622

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  58. An efficient fluorescence resonance energy transfer (FRET) between Pyrene and Perylene assembled in a DNA duplex and its potential for discriminating single-base changes

    Kashida H., Takatsu T., Sekiguchi K., Asanuma H.

    Chemistry - A European Journal   16 巻 ( 8 ) 頁: 2479 - 2486   2010年2月

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    記述言語:日本語   出版者・発行元:Chemistry - A European Journal  

    To increase the apparent Stokes' shift of perylene, pyrene (donor) and perylene (acceptor) were assembled in a DNA duplex to achieve the efficient fluorescence resonance energy transfer (FRET) from pyrene to perylene. Multiple donors were introduced in the vicinity of acceptors through d-threoninol and natural base pairs were inserted between the dyes. Accordingly, donors and acceptors could be accumulated inside the DNA without forming an undesired excimer/ exciplex. When two pyrene moieties were located in proximity to one perylene with one base pair inserted between them, efficient FRET occurred within the duplex. Thus, strong emission at 460 nm was observed from perylene when excited at 345 nm at which pyrene has its absorption. The apparent Stokes' shift became as large as 115 nm with a high apparent FRET efficiency (Fφ1). However, the introduction of more than two pyrenes did not enhance the fluorescence intensity of perylene, due to the short Fçrster radius (R0) of the donor pyrene. Next, this FRET system was used to enlarge the Stokes' shift of the DNA probe, which can discriminate a one-base deletion mutant from wild type with a model system by incorporation of multiple donors into DNA. Two perylene moieties were tethered to the DNA on both sides of the intervening base, and two pyrenes were further inserted in the vicinity of the perylenes as an antenna. Hybridization of this FRET probe with a fully matched DNA allowed monomer emission of perylene when the pyrenes were excited. In contrast, excimer emission was generated by hybridization with a one-base deletion mutant. Thus, the apparent Stokes' shift was enhanced without loss of efficiency in the detection of the deletion mutant. © 2010 Wiley-VCH Verlag GmbH&Co. KGaA, Weinheim.

    DOI: 10.1002/chem.200902078

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  59. Effect of the ortho modification of azobenzene on the photoregulatory efficiency of DNA hybridization and the thermal stability of its eis form

    Nishioka H., Liang X., Asanuma H.

    Chemistry - A European Journal   16 巻 ( 7 ) 頁: 2054 - 2062   2010年2月

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    記述言語:日本語   出版者・発行元:Chemistry - A European Journal  

    We synthesized various azobenzenes methylated at their ortho positions with respect to the azo bond for more effective photoregulation of DNA hybridization. Photoregulatory efficiency, evaluated from the change of T m (ΔTm) induced by trans-cis isomerization, was significantly improved for all ortho-modified azobenzenes compared with non-modified azobenzene due to the more stabilized trans form and the more destabilized cis form. Among the synthesized azobenzenes, 4-carboxy-2',6'- dimethylazobenzene (2',6'-Me-Azo), in which two ortho positions of the distal benzene ring with respect to carboxyl group were methylated, exhibited the largest ΔTm, whereas the newly synthesized 2,6-Me-Azo (4-carboxy-2,6- dimethylazobenzene), which possesses two methyl groups on the two ortho positions of the other benzene ring, showed moderate improvement of ΔTm. Both NMR spectroscopic analysis and computer modeling revealed that the two methyl groups on 2',6'-Me-Azo were located near the imino protons of adjacent base pairs; these stabilized the DNA duplex by stacking interactions in the trans form and destabilized the DNA duplex by steric hindrance in the cis form. In addition, the thermal stability of cis-2',6'-Me-Azo was also greatly improved, but not that of cis2,6-Me-Azo. Solvent effects on the half-life of the cis form demonstrated that cis-to-trans isomerization of all the modified azobenzenes proceeded through an inversion route. Improved thermal stability of 2',6'-Me-Azo but not 2,6-Me-Azo in the eis form was attributed to the retardation of the inversion process due to steric hindrance between lone pair electrons of the π orbital of the nitrogen atom and the methyl group on the distal benzene ring. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.

    DOI: 10.1002/chem.200902789

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  60. Femtosecond photoisomerization study on azobenzene-derivative bound by DNA

    Chen T., Igarashi K., Yamaguchi A., Nakagawa N., Yamane K., Fujii T., Asanuma H., Yamashita M.

    Optics InfoBase Conference Papers     2010年

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    記述言語:日本語   出版者・発行元:Optics InfoBase Conference Papers  

    First observation of femtosecond absorbance change in azobenzene-derivative (AzD) bound by double-strand DNA, that by single-strand DNA and Azd shows trans-to-cis photoisomerization rate per pulse in the former is much lower than in the latter. © OSA / UP 2010.

    DOI: 10.1364/up.2010.me6

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  61. Analysis of coherent heteroclustering of different dyes by Use of threoninol nucleotides for comparison with the molecular exciton theory

    Fujii T., Kashida H., Asanuma H.

    Chemistry - A European Journal   15 巻 ( 39 ) 頁: 10092 - 10102   2009年10月

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    記述言語:日本語   出版者・発行元:Chemistry - A European Journal  

    To test the molecular exciton theory for heterodimeric chromophores, various heterodimers and clusters, in which two different dyes were stacked alternately, were prepared by hybridizing two oligodeoxyribonucleotides (ODNs), each of which tethered a different dye on D-threoninol at the center of the strand. NMR analyses revealed that two different dyes from each strand were stacked antiparallel to each other in the duplex, and were located adjacent to the 5'-side of a natural nucleobase. The spectroscopic behavior of these heterodimers was systematically examined as a function of the difference in the wavelength of the dye absorption maxima (Δλmax). We found that the absorption spectrum of the heterodimer was significantly different from that of the simple sum of each monomeric dye in the single Strand. When azobenzene and Methyl Red, which have λmax at 336 and 480 nm, respectively, in the single strand (Δλmax= 144nm), were assembled on ODNs, the band derived from azobenzene exhibited a small hyperchromism, whereas the band from Methyl Red showed hypochromism and both bands shifted to a longer wavelength (bathochromism). These hyper- and hypochromisms were further enhanced in a heterodimer derived from 4'-methylthioazobenzene and Methyl Red, which had a much smaller Δλmax (82 nm; λmax = 398 and 480 nm in the single-strand, respectively). With a combination of 4'-dimethyl-amino-2- nitroazobenzene and Methyl Red, which had an even smaller Δλ max (33 nm), a single sharp absorption band that was apparently different from the sum of the single-stranded spectra was observed. These changes in the intensity of the absorption band could be explained by the molecular exciton theory, which has been mainly applied to the spectral behavior of H- and/or Jaggregates composed of homo dyes. However, the bathochromic band shifts observed at shorter wavelengths did not agree with the hypsochromism predicted by the theory. Thus, these data experimentally verify the molecular exciton theory of heterodimerization. This coherent coupling among the heterodimers could also partly explain the bathochromicity and hypochromicity that were observed when the dyes were intercalated into the duplex. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.

    DOI: 10.1002/chem.200900962

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  62. In-stem molecular beacon containing a pseudo base pair of threoninol nucleotides for the removal of background emission

    Kashida H., Takatsu T., Fujii T., Sekiguchi K., Liang X., Niwa K., Takase T., Yoshida Y., Asanuma H.

    Angewandte Chemie - International Edition   48 巻 ( 38 ) 頁: 7044 - 7047   2009年9月

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    記述言語:日本語   出版者・発行元:Angewandte Chemie - International Edition  

    Off means off: An in-stem molecular beacon in which Dthreoninol units tether perylene and anthraquinone in the stem region effectively detected target sequences and was able to discriminate a one-base-deletion mutant from the wild-type (full-match) sequence without background emission (see picture). © 2009 Wiley-VCH Verlag GmbH & Co. KGaA.

    DOI: 10.1002/anie.200902367

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  63. A supra-photoswitch involving sandwiched DNA base Pairs and azobenzenes for light-driven nanostructures and nanodevices

    Liang X., Mochizuki T., Asanuma H.

    Small   5 巻 ( 15 ) 頁: 1761 - 1768   2009年8月

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    記述言語:日本語   出版者・発行元:Small  

    A supra-photoswitch is designed for complete ON/OFF switching of DNA hybridization by light irradiation for the purpose of using DNA as a material for building nanostructures. Azobenzenes, attached to D-threoninols that function as scaffolds, are introduced into each DNA strand after every two natural nucleotides (in the form (NNX)n whereNandXrepresent the natural nucleotide and the azobenzene moiety, respectively). Hybridization of these two modified strands forms a supra-photoswitch consisting of alternating natural base pairs and azobenzene moieties. In this newly designed sequence, each base pair is sandwiched between two azobenzene moieties and all the azobenzene moieties are separated by base pairs. When the duplex is irradiated by visible light, the azobenzene moieties take the trans form and this duplex is surprisingly stable compared to the corresponding native duplex composed of only natural oligonucleotides. On the other hand, when the azobenzene moieties are isomerized to the cis form by UV light irradiation, the duplex is completely dissociated. Based on this design, a DNA hairpin structure is synthesized that should be closed by visible light irradiation and opened by UV light irradiation at the level of a single molecule. Indeed, perfect ON/OFF photoregulation is attained. This is a promising strategy for the design of supra-photoswitches such as photoresponsive sticky ends on DNA nanodevices and other nanostructures. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

    DOI: 10.1002/smll.200900223

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  64. Positively charged base surrogate for highly stable "base pairing" through electrostatic and stacking interactions

    Kashida H., Ito H., Fujii T., Hayashi T., Asanuma H.

    Journal of the American Chemical Society   131 巻 ( 29 ) 頁: 9928 - 9930   2009年7月

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    記述言語:日本語   出版者・発行元:Journal of the American Chemical Society  

    (Figure Presented) "Base pairs" of cationic dyes (p-methylstilbazole) were incorporated into oligodeoxyribonucleotides (ODNs). This "base pair" greatly stabilized the duplex through electrostatic and stacking interactions. The melting temperature of modified ODN was higher than those of neutral dyes and native base pairs. Further stabilization of the duplex was observed when the number of cationic dyes increased. © 2009 American Chemical Society.

    DOI: 10.1021/ja9013002

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  65. Modulation of pK<inf>a</inf> of Brooker's merocyanine by DNA hybridization

    Kashida H., Sano K., Kara Y., Asanuma H.

    Bioconjugate Chemistry   20 巻 ( 2 ) 頁: 258 - 265   2009年2月

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    記述言語:日本語   出版者・発行元:Bioconjugate Chemistry  

    Brocker's merocyanine (BM), which changes its emission and absorption maxima upon protonation, was introduced into oligodeoxyribonucleotide (ODN) via D-threoninol by postsynthetic modification on a CPG (controlled-pore glass) support. The pKa of BM in the modified ODN increased from 9.5 to 10.1 upon hybridization. As a result, absorption maxima shifted from 492 to 432 nm at pH 10.0 by the presence of its complementary strand. This spectral shift was sufficiently large so that DNA hybridization could easily be discriminated even by the naked eye; the color of the solution changed from orange to yellow upon hybridization. In addition, the fluorescence emission was strongly quenched upon hybridization, demonstrating that this probe can also detect the target DNA by the fluorescence change. Ratiometric detection of hybridization was also possible by simultaneous excitation of both protonated and deprotonated BMs. Furthermore, we could also modulate its pKa by the antiparallel stacking of two BM molecules in the duplex; the pKa of BM decreased from 10.1 to 9.7 by the stacking of two BMs in an antiparallel manner. Thus, control of the microenvironment around the BM molecule allowed modulation of its pKa, which is applicable to the sequence-specific recognition of target DNA. © 2009 American Chemical Society.

    DOI: 10.1021/bc800335h

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  66. Rational design of functional DNA with a Non-Ribose acyclic scaffold

    Kashida H., Liang X., Asanuma H.

    Current Organic Chemistry   13 巻 ( 11 ) 頁: 1065 - 1084   2009年

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    記述言語:日本語   出版者・発行元:Current Organic Chemistry  

    The growing field of DNA technology requires new modified DNAs that can perform advanced functions. No matter how we optimize the length and sequence of DNA using only the four naturally occurring nucleotides, potential performance is limited. In this review, we describe a facile and effective method of rationally designing new functional DNA by focusing on acyclic scaffolds, especially threoninols, which are utilized to incorporate functional molecules into DNA. Wedge-type insertion of a functional molecule with a planar structure of proper size in D-threoninol to DNA does not destabilize the duplex, although the backbone structure is changed. Rather, intercalation offsets such distortions and significantly raises the melting temperature of the DNA duplex. Based on the wedge-type insertion, photoresponsive DNA (tethering azobenzenes) and fluorescent probes that can detect single nucleotide polymorphisms (SNPs) and insertion/deletion (indel) polymorphisms have been designed. Furthermore, a variety of molecular clusters of dyes have also been prepared from acyclic scaffolds tethering dyes. © 2009 Bentham Science Publishers Ltd.

    DOI: 10.2174/138527209788680736

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  67. 名古屋コンファレンス報告

    浅沼 浩之, 早川 芳宏

    化学と工業 = Chemistry and chemical industry   60 巻 ( 4 )   2007年4月

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    記述言語:日本語  

  68. 2′,6′-Dimethylazobenzene as an efficient and thermo-stable photo-regulator for the photoregulation of DNA hybridization

    Nishioka H., Liang X., Kashida H., Asanuma H.

    Chemical Communications   ( 42 ) 頁: 4354 - 4356   2007年

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    記述言語:日本語   出版者・発行元:Chemical Communications  

    The introduction of methyl groups into two ortho positions (2′ and 6′ positions) of the same benzene ring in an azobenzene remarkably raised both its photoregulation ability and the thermal stability of the cis-form. © The Royal Society of Chemistry.

    DOI: 10.1039/b708952j

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  69. Unexpected efficient ab initio DNA synthesis at low temperature by using thermophilic DNA polymerase.

    Liang X., Kato T., Asanuma H.

    Nucleic acids symposium series (2004)   ( 51 ) 頁: 351 - 352   2007年

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    記述言語:日本語   出版者・発行元:Nucleic acids symposium series (2004)  

    DNA was synthesized in the absence of DNA or RNA as template (or primer) from dNTPs at relatively low temperatures (25 approximately 50 degrees C) by thermophilic Vent DNA polymerase whose proper reaction temperature for primer extension is 70 approximately 80 degrees C. Unexpectedly, the ab initio DNA synthesis was even more efficient at 50 degrees C as compared with that at 70 degrees C. Interestingly, the ab initio DNA synthesis by Vent (exo), a mutant version of Vent DNA polymerase lacking of 3' --> 5' exonuclease activity, became much less efficient, and it could only carry out ab initio DNA synthesis after a long incubation time. This remarkable difference between Vent and Vent (exo) indicates that the exonuclease activity domain of Vent may play an important role at the initiation step of ab initio DNA synthesis.

    DOI: 10.1093/nass/nrm176

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  70. Effective photoregulation of DNA hybridization by the introduction methyl group into azobenzene

    Nishioka H., Kashida H., Komiyama M., Asanuma H.

    Polymer Preprints, Japan   55 巻 ( 1 )   2006年10月

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    記述言語:日本語   出版者・発行元:Polymer Preprints, Japan  

    We have synthesized azobenzene-tethered DNA and have successfully photoregulated various DNA functions. In the present study, we synthesized various azobenzenes substituted with methyl group for still more effective photoregulation, as shown in Fig. 1. The melting temperatures of 1a/S 0 duplex are listed in Table 1. When an azobenzene took trans-form, mono-substituted one at ortho position exhibited larger Tm compared with other mono-substituted ones. In contrast, Tm of ortho-substituted azobenzene was the smallest in Tm-form. As a result, change of T m (ΔTm) induced by trans-cis isomerization was as large as 10.6°C, which was 4.9°C higher than previous non-substituted azobenzene (Azo: 5.7°C). Quite interestingly, di-substituted azobenzene at both ortho positions (2,6-Azo) exhibited even larger ΔTm (14.6°C). Thus, we succeeded in the efficient photoregulation of DNA hybridization by the modification of azobenzene at ortho position.

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  71. Synthesis of peptide nucleic acid conjugated with carbohydrate

    Noguchi A., Yamada Y., Nishida Y., Asanuma H.

    Polymer Preprints, Japan   55 巻 ( 1 )   2006年10月

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    記述言語:日本語   出版者・発行元:Polymer Preprints, Japan  

    Peptide nucleic acid (PNA) is composed of non-charged N-2-aminoethylglycine as a scaffold, and thus it strongly hybridizes with natural DNA. But due to its high hydrophobic property, large PNA oligomers prefer to make aggregates rather than form duplex with DNA. Therefore, incorporation of functional molecules such as intercalators is difficult because such modification should raise the hydrophobicity of PNA. In order to modify the PNA without increasing its hydrophobicity, we designed a new monomer involving carbohydrate for the introduction of PNA oligomer. Here, we successfully synthesized a lysine monomer tethering mannose unit according to the Scheme 1. In the present paper, effect of mannose on the stability of PNA/DNA duplex is examined.

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  72. Synthesis of fluorescent DNA probe by use of dye polarization

    Sano K., Tanaka M., Kashida H., Komiyama M., Asanuma H.

    Polymer Preprints, Japan   55 巻 ( 1 )   2006年10月

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    記述言語:日本語   出版者・発行元:Polymer Preprints, Japan  

    DNA duplex involves hydrophobic inside in which water molecules are excluded, and its exterior is surrounded with the anions of phosphodiester linkages. Since the inside is not shielded by water molecules, strong electric field is formed by the phosphodiester anion and thus the molecules bound to the duplex should be easily polarized. In the present study, we tethered Dansyl moiety as a fluorophore through amide bond to DNA (Dco in Scheme 1), and fluorescence change by duplex formation was investigated. Single-stranded S showed fluorescence emission at around 550 nm (Blue line in Fig.1). When it was hybridized with native S0, small but distinct decrease in the fluorescence by the phosphodiester anion was observed (Red line). On the contrary, hybridization with M56 (Green line), in which two phosphodiesters nearest to Dansyl moiety are replaced with non-charged methyl phosphonate, slightly increased the fluorescence.

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  73. Stabilization of DNA duplex by introducing intercalators into DNA through D-threoninol

    Kashida H., Komiyama M., Asanuma H.

    Polymer Preprints, Japan   55 巻 ( 1 )   2006年10月

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    記述言語:日本語   出版者・発行元:Polymer Preprints, Japan  

    We have introduced various dyes into DNA through D-threoninol and successfully functionalized DNA. However, little has been known on the relationship between the structure of intercalators and the stability of duplex. In this study, various intercalators (1 to 7 as shown in Scheme 1) were incorporated into DNA and the melting temperatures (TmS) of duplexes were measured. As shown in Table 1, azobenzene (1) stabilized duplex strongly compared with native duplex. However, larger molecules (2 and 3) and a smaller molecule (4) did not stabilize duplex. Therefore, molecular size is one of the most significant factors for the duplex stability.

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  74. Recognition of bioactive oligopeptide by imprinted cyclodextrin polymer in water

    Sumaoka J., Shirasaka K., Katayama M., Song S., Nagaoka S., Asanuma H., Komiyama M.

    Polymer Preprints, Japan   55 巻 ( 1 )   2006年10月

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    記述言語:日本語   出版者・発行元:Polymer Preprints, Japan  

    We prepared molecularly imprinted polymers of β-cyclodextrin using three bioactive oligopeptides as templates; (a) angiotensin I (10-mer), (b) γ-endorphin (17-mer), and (c) leucine-enkephalin (Leu-Enk: 6-mer). With both angiotensin I and Leu-Enk as the templates, highly precise imprinting effects were observed. The present finding strongly indicates that we can prepare artificial antibodies by imprinting of CyD to a conformationally restrained part of a protein.

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  75. Preparation of new dye supramolecule by the spontaneous aggregation

    Yamada A., Kashida H., Komiyama M., Asanuma H.

    Polymer Preprints, Japan   55 巻 ( 1 )   2006年10月

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    記述言語:日本語   出版者・発行元:Polymer Preprints, Japan  

    In the previous study, we introduced cationic dyes (Naphthyl Reds) and spacers alternately into the middle of 12mer oligonucleotides and prepared dye cluster by using DNA hybridization. The melting temperature (Tm) of this duplex was much higher than native DNA due to electrostatic and stacking interaction. Therefore, there is a possibility of forming dye cluster without the aid of natural bases. In this study, we decreased the number of natural bases at the terminal from 12 to 6 and 2 (see Scheme 1) and investigated the formation of dye cluster. Circular dichroism of 3C/3D and 3G/3G showed very strong symmetrical positive and negative Cotton effect at around 500 nm, suggesting the formation of dye cluster. The melting temperature (Tm) determined from the change of CD at 536 nm of 3C/3D was about 30°C, which almost coincided with the Tm from absorbance change at 260nm. Similarly, melting curve of 3C/3D of CD intensity at 536 nm also showed loose sigmoid, suggesting that 3G/3G also forms dye cluster.

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  76. Preparation of active DNA enzymes by introduction of intercalator

    Hayashi H., Zhao J., Komiyama M., Liang X., Asanuma H.

    Polymer Preprints, Japan   55 巻 ( 1 )   2006年10月

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    記述言語:日本語   出版者・発行元:Polymer Preprints, Japan  

    The RNA cleaving 10-23 DNA enzyme, which is expected to be used as an antisense agent to suppress gene activity, shows dependence of activity on the sequence at the active site. We already showed that the cleavage efficiency of 10-23 DNA enzyme with lower activity was improved by attaching an azobenzene residue close to the active site. In this study, we introduced several planar molecules instead of azobenzene as intercalators and checked their activation (Fig. 1). We found that structures of introduced intercalators and linkers greatly influenced the cleavage activity. When anthraquinone was used as the intercalator, catalytic activation was increased more than six folds in comparison with the native DNA enzyme (Table 1). Our finding is helpful for understanding the mechanism of cleavage by DNA enzymes.

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  77. Importance of the position of vinyl group on β-cyclodextrin for the effective imprinting of amino acid derivatives and oligopeptides in water

    Osawa T., Shirasaka K., Matsui T., Yoshihara S., Akiyama T., Hishiya T., Asanuma H., Komiyama M.

    Macromolecules   39 巻 ( 7 ) 頁: 2460 - 2466   2006年4月

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    記述言語:日本語   出版者・発行元:Macromolecules  

    Two kinds of vinyl monomers of β-cyclodextrin (β-CyD) that tether a vinyl group on either wider rim of the truncated cone or its smaller rim were synthesized and applied to the imprinting toward amino acid derivatives and oligopeptides in water. Mono-3-(N-acrylamido)-β-deoxy-altro-β- cyclodextrin (3-AAm-CyD) showed a remarkable imprinting effect for the enantioselective recognition of protected amino acids such as N-benzyloxycarbonyltyrosine (Z-Tyr). However, the imprinted polymer from mono-6-(N-acrylamido)-6-deoxy-βcyclodextrin (6-AAm-CyD) hardly showed enantioselectivity. According to NOESY analysis on the preorganized β-CyD/Z-Tyr complex in D2O, the aromatic moieties of Z-Tyr were included into the cavity of β-CyD from its wider rim. Since the vinyl group of 3-AAm-CyD protruded toward the template and was polymerized there, the detailed shape of the template was precisely copied on the polymer by the imprinting. In the case of 6-AAm-CyD, however, the shape of template could not be well transcribed because its vinyl group was located at opposite side of the cavity, and thus copolymerization occurred far from the template molecule. On the other hand, the imprinted polymers from both β-CyD vinyl monomers were effective for the recognition of sequences of tetrapeptides composed of two glycines and two phenylalanines, although the selectivity itself was not remarkable. In these polymers, even the β-CyD residues of 6-AAm-CyD were immobilized complementarily to the phenyl rings and bound them. © 2006 American Chemical Society.

    DOI: 10.1021/ma060064f

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  78. Photo- and thermoregulation of DNA nanomachines

    Takahashi K., Yaegashi S., Asanuma H., Hagiya M.

    Lecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics)   3892 LNCS 巻   頁: 336 - 346   2006年

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    記述言語:日本語   出版者・発行元:Lecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics)  

    We have been investigating DNA state machines, especially those based on the opening of hairpin molecules in which state transitions are realized as hairpin loops are opened by molecules called openers. This paper introduces photo- and thermoregulation of such hairpin-based DNA machines, in which the openers become active by sensing external signals in the form of light or heat. We conducted fluorescence experiments and show that photo- and thermoregulation is possible. In the experiments, the openers become active when they are irradiated by UV light or when they receive heat as external input. For photoregulation, we use azobenzene-bearing oligonucleotides developed by the third author. © Springer-Verlag Berlin Heidelberg 2006.

    DOI: 10.1007/11753681_26

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  79. Functionalization of DMA by using DNA-pyrene conjugates

    Kashida H., Komiyama M., Asanuma H.

    Polymer Preprints, Japan   54 巻 ( 2 ) 頁: 5092 - 5093   2005年12月

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    記述言語:日本語   出版者・発行元:Polymer Preprints, Japan  

    We have successfully introduced functional molecules into DNA and prepared various aggregates by intercalation between base-pairs. Here, we synthesized DNA-pyrene conjugates through D-threoninol and investigated their properties. Excimer emission of pyrene was not observed when two pyrene moieties were intervened by one base-pair. On the other hand, strong excimer emission was observed at around 480 nm with complementary strand which lacks one base. Sequence dependence and linker modification are also investigated.

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  80. Photo- and Thermoregulation of DNA Nanomachines

    浅沼 浩之, 萩谷 昌己

    DNA11, Eleventh International Meeting on DNA Based Computers, Preliminary Proceedings     頁: 147 - 156   2005年

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  81. バイオナノデバイスとしての人工DNA 査読有り

    浅沼 浩之

    化学と教育 53     頁: 12 - 15   2005年

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  82. NMR study on the photoresponsive DNA tethering an azobenzene. Assignment of the absolute configuration of two diastereomers and structure determination of their duplexes in the trans-form.

    Liang X, Asanuma H, Kashida H, Takasu A, Sakamoto T, Kawai G, Komiyama M

    Journal of the American Chemical Society   125 巻 ( 52 ) 頁: 16408 - 15   2003年12月

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    記述言語:英語  

    DOI: 10.1021/ja037248j

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  83. DNA-Naphthyl Red conjugate as a visualizing probe of DNA hybridization.

    Asanuma H, Kashida H, Liang X, Komiyama M

    Chemical communications (Cambridge, England)   ( 13 ) 頁: 1536 - 7   2003年7月

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    記述言語:英語  

    DOI: 10.1039/b302875e

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  84. DNA-dye conjugates for controllable H aggregation(1).

    Asanuma H, Shirasuka K, Takarada T, Kashida H, Komiyama M

    Journal of the American Chemical Society   125 巻 ( 8 ) 頁: 2217 - 23   2003年2月

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    記述言語:英語  

    DOI: 10.1021/ja021153k

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  85. Development of a probe DNA which accompanies color change on hybridization.

    Kashida H, Asanuma H, Komiyama M

    Nucleic acids research. Supplement (2001)   ( 3 ) 頁: 143 - 4   2003年

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    記述言語:英語  

    DOI: 10.1093/nass/3.1.143

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  86. Photo-regulation of DNA function by azobenzene-tethered oligonucleotides.

    Asanuma H, Matsunaga D, Liu M, Liang X, Jhao J, Komiyama M

    Nucleic acids research. Supplement (2001)   ( 3 ) 頁: 117 - 8   2003年

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    記述言語:英語  

    DOI: 10.1093/nass/3.1.117

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  87. Effective photo-regulation of transcription reaction by SP6 RNA polymerase with modified DNA tethering multiple azobenzenes.

    Liu M, Asanuma H, Komiyama M

    Nucleic acids research. Supplement (2001)   ( 3 ) 頁: 265 - 6   2003年

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    記述言語:英語  

    DOI: 10.1093/nass/3.1.265

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  88. Photoregulation of the transcription reaction of T7 RNA polymerase by tethering an azobenzene to the promoter.

    Asanuma H, Tamaru D, Yamazawa A, Liu M, Komiyama M

    Chembiochem : a European journal of chemical biology   3 巻 ( 8 ) 頁: 786 - 9   2002年8月

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  89. Photoregulation of DNA triplex formation by azobenzene.

    Liang X, Asanuma H, Komiyama M

    Journal of the American Chemical Society   124 巻 ( 9 ) 頁: 1877 - 83   2002年3月

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    記述言語:英語  

    DOI: 10.1021/ja011988f

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  90. Spectroscopic anatomy of molecular-imprinting of cyclodextrin. Evidence for preferential formation of ordered cyclodextrin assemblies.

    Hishiya T, Asanuma H, Komiyama M

    Journal of the American Chemical Society   124 巻 ( 4 ) 頁: 570 - 5   2002年1月

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    記述言語:英語  

    DOI: 10.1021/ja011305w

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  91. Photo-regulation of transcription by RNA polymerase with azobenzene-tethered promoter.

    Asanuma H, Liu M, Tamaru D, Liang X, Komiyama M

    Nucleic acids research. Supplement (2001)   ( 2 ) 頁: 75 - 6   2002年

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    記述言語:英語  

    DOI: 10.1093/nass/2.1.75

    PubMed

  92. Enantioselective Incorporation of Azobenzenes into Oligodeoxyribonucleotide for Effective Photoregulation of Duplex Formation This work was partially supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (Molecular Synchronization for Design of New Materials System). The support by the Grant from "Research for the Future" Program of the Japan Society for the Promotion of Science (JSPS-RFTF97I00301) is also acknowledged.

    Asanuma H, Takarada T, Yoshida T, Tamaru D, Liang X, Komiyama M

    Angewandte Chemie (International ed. in English)   40 巻 ( 14 ) 頁: 2671 - 2673   2001年7月

     詳細を見る

    記述言語:英語  

    PubMed

  93. Enantioselective Incorporation of Azobenzenes into Oligodeoxyribonucleotide for Effective Photoregulation of Duplex Formation.

    Asanuma H, Takarada T, Yoshida T, Tamaru D, Liang X, Komiyama M

    Angewandte Chemie (International ed. in English)   40 巻 ( 14 ) 頁: 2671 - 2673   2001年7月

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  94. Acceleration of influenza virus clearance by Th1 cells in the nasal site of mice immunized intranasally with adjuvant-combined recombinant nucleoprotein

    Tamura S.I., Miyata K., Matsuo K., Asanuma H., Takahashi H., Nakajima K., Suzuki Y., Aizawa C., Kurata T.

    Journal of Immunology   156 巻 ( 10 ) 頁: 3892 - 3900   1996年5月

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    記述言語:日本語   出版者・発行元:Journal of Immunology  

    The protective roles of influenza viral nucleoprotein (NP), together with the cellular mechanism of the protection in the nasal site, were examined in BALB/c mice immunized intranasally with an adjuvant (cholera toxin B subunit containing 0.2% of the whole toxin)-combined A or B virus recombinant NP. The NP-immune mice, when challenged intranasally with a sublethal dose of the virus 3 wk after immunization, had accelerated virus clearance from the nasal site in both an influenza type-specific and a nonspecific manner, as shown by the protection from high morbidity from the second day after challenge. Both type-specific and nonspecific acceleration of recovery was confirmed by the increased survival rate after challenge with a lethal dose of virus in mice immunized and boosted with adjuvant-combined NP. The acceleration of nasal virus clearance was accompanied with acceleration of type-specific systemic delayed-type hypersensitivity (DTH) and with IFN-γ production by nasal lymphocytes. The nasal lymphocytes from the immunized and challenged mice generated a significantly high level of DTH when transferred locally, but no class I MHC-restricted CTL response. Moreover, nasal CD4+ T cells, induced by NP immunization and increased in number by the subsequent challenge, were involved in the accelerated IFN-γ production. These results suggest that nasal Th1 cells, capable of producing IFN-γ and mediating DTH, are involved in the type-specific acceleration of recovery from influenza after challenge in mice immunized intranasally with adjuvant-combined NP, although the nonspecific mechanism of accelerated recovery remains to be solved.

    Scopus

  95. Signal Amplification Circuit Composed of Serinol Nucleic Acid for RNA Detection 査読有り

    Chen, Y.; Murayama, K.; Asanuma, H.

    Chemistry Letters   51 巻   頁: 330 - 333   2022年2月

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    担当区分:最終著者, 責任著者   記述言語:英語  

    DOI: 10.1246/cl.210813

  96. Renewable DNA proportional-integral controller with photoresponsive molecules 招待有り 査読有り

    Tamba, M.; Murayama, K.; Asanuma, H.; Nakakuki, T.

    Micromachines   13 巻   頁: 193   2022年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: doi.org/10.3390/mi13020193

  97. Unexpected Dissociation of Photoresponsive UV-ON DNA Carrying p-tert-Butyl Azobenzene under UV Light Irradiation 査読有り

    Ishii, S.; Murayama, K.; Sada, K.; Asanuma, H.; Kakugo, A.

    Chemistry Letters   51 巻   頁: 292 - 295   2022年1月

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    担当区分:責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1246/cl.210788

  98. 光駆動型DNAナノマシン 招待有り 査読有り

    浅沼浩之, 村山恵司, 神谷由紀子, 樫田 啓

    高分子   70 巻   頁: 550 - 552   2021年10月

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    担当区分:筆頭著者, 責任著者   記述言語:日本語   掲載種別:研究論文(学術雑誌)  

  99. Microheater-integrated zinc oxide nanowire microfluidic device for hybridization-based detection of target single-stranded DNA 査読有り

    Nanotechnology     2021年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  100. Development of pre-miRNAs modification with FRET dye pair for intracellular visualization of processing intermediates generated in cells 査読有り

    Kamiya, Y.; Kamimoto, H.; Zhu, H. ; Asanuma, H.

    Sensors   21 巻   頁: 1785   2021年3月

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    担当区分:最終著者, 責任著者   記述言語:英語  

    DOI: doi.org/10.3390/s21051785

  101. Pyrene modified serinol nucleic acid nanostructure converts chirality of threoninol nucleic acids into circularly polarized luminescence signals 招待有り 査読有り

    Kashida, H.; Nishikawa, K.; Ito Y.; Murayama, K.; Hayashi, I.; Kakuta, T.; Ogoshi, T.; Asanuma, H.

        2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/chem.202102333

  102. Microheater-integrated zinc oxide nanowire microfluidic device for hybridization-based detection of target single-stranded DNA 査読有り

    Takahashi, H.; Yasui, T.; Kashida, H.; Makino, K.; Shinjo, K.; Liu, Q.; Shimada, T.; Rahong, S.; Kaji, N.; Asanuma, H.; Baba, Y.

    Nanotechnology   32 巻   頁: 255301   2021年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1088/1361-6528/abef2c

  103. 核酸医薬の創製に向けた人工核酸の開発 招待有り 査読有り

    樫田啓、村山恵司、浅沼浩之

    バイオマテリアル   38 巻 ( 2 ) 頁: 124 - 129   2020年4月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

  104. The Use of Serinol Nucleic Acids as Ultrasensitive Molecular Beacons 招待有り 査読有り

    Murayama Keiji, Kashida Hiromu, Asanuma Hiroyuki

    NON-NATURAL NUCLEIC ACIDS: METHODS AND PROTOCOLS   1973 巻   頁: 261-279   2019年

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/978-1-4939-9216-4_17

    Web of Science

  105. Development of Visible-Light-Responsive RNA Scissors Based on a 10-23 DNAzyme 査読有り

    Kamiya Yukiko, Arimura Yu, Ooi Hideaki, Kato Kenjiro, Liang Xing-Guo, Asanuma Hiroyuki

    CHEMBIOCHEM   19 巻 ( 12 ) 頁: 1305 - 1311   2018年6月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/cbic.201800020

    Web of Science

  106. DNA Microcapsule for Photo-Triggered Drug Release Systems

    Kamiya Yukiko, Yamada Yoshinobu, Muro Takahiro, Matsuura Kazunori, Asanuma Hiroyuki

    CHEMMEDCHEM   12 巻 ( 24 ) 頁: 2016-2021   2017年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/cmdc.201700512

    Web of Science

  107. Introduction of 2,6-Diaminopurines into Serinol Nucleic Acid Improves Anti-miRNA Performance 査読有り

    Kamiya Yukiko, Donoshita Yuka, Kamimoto Hiroshi, Murayama Keiji, Ariyoshi Jumpei, Asanuma Hiroyuki

    CHEMBIOCHEM   18 巻 ( 19 ) 頁: 1917-1922   2017年10月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/cbic.201700272

    Web of Science

  108. Chaperone-Polymer-Assisted, Photodriven DNA Strand Displacement

    Cheng Bohao, Kashida Hiromu, Shimada Naohiko, Maruyama Atsushi, Asanuma Hiroyuki

    CHEMBIOCHEM   18 巻 ( 16 ) 頁: 1568-1572   2017年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/cbic.201700202

    Web of Science

  109. D-aTNA Circuit Orthogonal to DNA Can Be Operated by RNA Input via SNA

    Murayama Keiji, Nagao Ryuya, Asanuma Hiroyuki

    CHEMISTRYSELECT   2 巻 ( 20 ) 頁: 5624-5627   2017年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/slct.201701126

    Web of Science

  110. Antisense oligonucleotide modified with serinol nucleic acid (SNA) induces exon skipping in mdx myotubes 査読有り

    Le, B. T.; Murayama, K.; Shabanpoor, F.; Asanuma, H.; Veedu, R. N.

    RSC Advances   7 巻   頁: 34049-34052   2017年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: DOI: 10.1039/c7ra06091b

  111. 光応答性DNAの開発―ナノ環境を汚染しない分子マシンへの応用― 招待有り

    浅沼浩之

    現代化学   556 巻 ( 7 ) 頁: 50-55   2017年7月

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    担当区分:筆頭著者   記述言語:日本語  

  112. DNA二重鎖の高い安定性と直交性をもつ疑似塩基対の開発 招待有り 査読有り

    樫田啓、浅沼浩之

    高分子論文集   74 巻 ( 4 ) 頁: 257-264   2017年7月

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    担当区分:筆頭著者   記述言語:日本語  

  113. Orientation-dependent FRET system reveals differences in structures and flexibilities of nicked and gapped DNA duplexes

    Kashida Hiromu, Kurihara Ayako, Kawai Hayato, Asanuma Hiroyuki

    NUCLEIC ACIDS RESEARCH   45 巻 ( 11 )   2017年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1093/nar/gkx200

    Web of Science

  114. Development of pseudo base-pairs on D-threoninol which exhibit various functions 招待有り 査読有り

    Kashida, H.; Asanuma, H.

    Bull. Chem. Soc. Jpn.   90 巻   頁: 475   2017年5月

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    担当区分:筆頭著者   記述言語:英語  

  115. Design of photo-functional oligonucleotide by "copolymerization" of natural nucleobases with base-surrogate prepared from acyclic scaffold 招待有り 査読有り

    Asanuma, H.; Murayama, K.; Kamiya, Y.; Kashida, H.

    Polymer J.   49 巻   頁: 279-289   2017年3月

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    担当区分:筆頭著者   記述言語:英語  

    DOI: doi:10.1038/pj.2016.120

  116. DNA二重鎖による色素会合体の精密位置配向制御と光化学への展開 招待有り 査読有り

    浅沼浩之、樫田啓

    固体物理   52 巻 ( 3 ) 頁: 149-158   2017年3月

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    担当区分:筆頭著者   記述言語:日本語  

  117. Effect of Methyl Group on Acyclic Serinol Scaffold for Tethering Dyes on the DNA Duplex Stability

    Murayama Keiji, Asanuma Hiroyuki

    CHEMBIOCHEM   18 巻 ( 1 ) 頁: 142-149   2017年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/cbic.201600558

    Web of Science

  118. Orientation-dependent FRET system reveals differences in structures and flexibilities of nicked and gapped DNA duplexes 査読有り

    Kashida, H.; Kurihara, A.; Kawai, H.; Asanuma, H.

    Nucleic Acids Res.   45 巻 ( 11 ) 頁: e105   2017年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  119. DNA二重鎖の高い安定性と直交性をもつ疑似塩基対の開発

    樫田 啓, 浅沼 浩之

    高分子論文集   74 巻 ( 4 ) 頁: 257 - 264   2017年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 高分子学会  

    天然核酸塩基は高い二重鎖安定性と直交性という化学的に魅力的な性質をもつ.この天然塩基対に代わる新たな塩基対を開発することができれば,天然がもちえない機能をもった核酸材料の開発が可能となる.筆者らは非環状骨格である<small>D</small>-threoninolに着目し,これを介した疑似塩基対の開発を行ってきた.また,疑似塩基対の化学構造が天然塩基対とまったく異なるにもかかわらず二重鎖の安定化と直交性という天然塩基対の化学的機能を模倣できることを明らかにした.本報ではとくに疑似塩基対の安定性に焦点を当て,筆者らの最近の研究成果と今後の展望について概説する.

    DOI: 10.1295/koron.2017-0009

    Web of Science

    Scopus

  120. Acyclic artificial nucleic acids with phosphodiester bonds exhibit unique functions 招待有り 査読有り

    H. Kashida, K. Murayama, H. Asanuma

    Polymer J.   48 巻   頁: 781-786   2016年

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    記述言語:英語  

  121. A stem-less probe using spontaneous pairing between Cy3 and quencher for RNA detection 招待有り 査読有り

    Kashida, H., Morimoto, K., Asanuma, H.

    Sci. Technol. Adv. Mater.   17 巻   頁: 267-273   2016年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  122. Faint electric treatment-induced rapid endosomal escape for efficient delivery of functional macromolecules into the cytoplasm 査読有り

    Hasan, M., Nishimoto, A., Ohgita, T., Hama, S., Kashida, H., Asanuma, H., Kogure, K.

    J. Control. Release   228 巻   頁: 20-25   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  123. Visible Light-Triggered Crosslinking of DNA Duplex by Reversible [2+2] Photocycloaddition of Styrylpyrene 査読有り

    Doi, T., Kawai, H., Murayama, K., Kashida, H., Asanuma, H.

    Chem. Eur. J.     頁: 0000-0000   2016年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 0000-0000

  124. Strand-invading linear probe combined with unmodified PNA 査読有り

    Aasanuma, H., Niwa, R., Akahane, M., Murayama, K., Kashida, H., Kamiya, Y.

    Bioorg. Med. Chem.     2016年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: DOI:10.1016/j.bmc.2016.06.055

  125. Dynamics of Inter-DNA Chain Interaction of Photoresponsive DNA 査読有り

    Nakasone, Y., Ooi, H., Kamiya, Y., Asanuma, H., Terazima, M.

    J. Am. Chem. Soc.     頁: 0000-0000   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  126. 天然に存在しない非環状構造の核酸アナログ-人工の非環状骨格で神の設計した環状骨格に挑む 招待有り

    村山恵司、浅沼浩之

    化学   71 巻 ( 3 ) 頁: 66-67   2016年

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    担当区分:筆頭著者   記述言語:日本語  

  127. Molecular design of Cy3 derivative for highly sensitive in-stem molecular beacon and its application to the wash-free FISH 査読有り

    H. Kashida, T. Osawa, K. Morimoto, Y. Kamiya, H. Asanuma

    Bioorg. Med. Chem.   23 巻   頁: 1758-1762   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  128. Efficiency of [2+2] photodimerization of various stilbene derivatives within the DNA duplex scaffold 査読有り

    T. Doi, H. Kashida, H. Asanuma

    Org. Biomol. Chem.   13 巻   頁: 4430-4437   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  129. Highly sensitive and robust linear probe for detection of mRNA in cells 査読有り

    H. Asanuma, M. Akahane, R. Niwa, H. Kashida, Y. Kamiya

    Angew. Chem. Int. Ed.   54 巻   頁: 4315-4319   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  130. Acyclic L-Threoninol Nucleic Acid (L-aTNA) with Suitable Structural Rigidity Cross-pairs with DNA and RNA 査読有り

    K. Murayama, H. Kashida, H. Asanuma,

    Chem. Commun.   51 巻   頁: 6500-6503   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  131. Reversible photoswitching of RNA hybridization at room temperature with an azobenzene C-nucleoside 査読有り

    T. Goldau, K. Murayama, C. Brieke, S. Steinwand, P. Mondal, M. Biswas, I. Burghardt, J. Wachtveitl, H. Asanuma, A. Heckel

    Chem. Eur. J.   21 巻   頁: 2845-2854   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  132. Synthetic gene involving azobenzene-tethered T7 promoter for the photocontrol of gene expression by visible light 査読有り

    Y. Kamiya, T. Takagi, H. Ooi, H. Ito, X.G. Liang, H. Asanuma,

    ACS Synth. Biol.   4 巻   頁: 365-370   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  133. Ultra-Sensitive Molecular Beacon Designed with Totally Serinol Nucleic Acid (SNA) for Monitoring mRNA in Cell 招待有り 査読有り

    K. Murayama, Y. Kamiya, H. Kashida, H. Asanuma

    ChemBioChem   16 巻   頁: 1298-1301   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: DOI: 10.1002/cbic.201500167

  134. 新規人工核酸SNAを用いたRNAイメージング 招待有り

    樫田 啓、村山恵司、浅沼浩之

    生体の科学   66 巻 ( 2 ) 頁: 145-150   2015年

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    担当区分:筆頭著者   記述言語:日本語  

  135. Reversible photo-switching of DNA fuction with azobenzene-tethered DNA 招待有り

    Y. Kamiya, H. Asanuma

    The Glen Report   27 巻   頁: 1-3   2015年

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    担当区分:筆頭著者   記述言語:英語  

  136. Pre-organized guide RNA in Cas9 complex is ready for selection of the target double-stranded DNA 招待有り 査読有り

    Y. Kamiya, H. Asanuma

    ChemBioChem   16 巻   頁: 2273-2275   2015年

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    担当区分:筆頭著者   記述言語:英語  

  137. Terminus-free siRNA prepared by photo-crosslinking activated via slicing by Ago2 査読有り

    Y. Kamiya, K. Iishiba,T. Doi, K. Tsuda, H. Kashida, H. Asanuma,

    Biomater. Sci.   3 巻   頁: 1534-1538   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  138. Azobenzene C-nucleosides for photo-controlled hybridization of DNA at room temperature 査読有り

    T. Goldau, K. Murayama, C. Brieke, H. Asanuma, A. Heckel

    Chem. Eur. J.   21 巻   頁: 17870-17876   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: DOI: 10.1002/chem.201503303

  139. Hetero-Selective DNA-like Duplex Stabilized by Donor-Acceptor Interaction 査読有り

    T. Doi, T. Sakakibara,H. Kashida, Y. Araki, T. Wada, H. Asanuma

    Chem. Eur. J.   21 巻   頁: 15974-15980   2015年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: DOI: 10.1002/chem.201502653

  140. DNA二重鎖を足場とした配向依存的FRET 招待有り

    浅沼浩之、栗原綾子、樫田啓

    光化学   45 巻   頁: 146-147   2014年12月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  141. Light-driven DNA nanomachine with a photoresponsive molecular engine 招待有り 査読有り

    Kamiya, Y.; Asanuma, H.

    Accounts of Chemical Research   47 巻   頁: 1663-1672   2014年

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    担当区分:筆頭著者   記述言語:英語  

  142. De Novo Design of Functional Oligonucleotides with Acyclic Scaffolds 招待有り 査読有り

    Asanuma, H.; Kashida, H.; Kamiya, Y.

    Chem. Rec.     2014年

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    担当区分:筆頭著者   記述言語:英語  

    DOI: DOI:10.1002/tcr.201402040

  143. Enhancement of stability and activity of siRNA by terminal substitution with Serinol Nucleic Acid (SNA) 査読有り

    Y. Kamiya, J. Takai, H. Ito, K. Murayama, H. Kashida, H. Asanuma.

    ChemBioChem   15 巻   頁: 2549-2555   2014年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  144. Highly specific DNA detection from massive background nucleic acids based on rolling circle amplification of target dsDNA 査読有り

    X. Wang, X. Yu, X. Wand, M. Suzuki, H. Asanuma, P. Dong, W. Wu, X.G. Liang,

    RSC Adv.   4 巻   頁: 38293-38299   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  145. Selective labeling of mature RISC using siRNA carrying fluorophore-quencher pair 査読有り

    Kamiya, Y.; Ito, A.; Ito, H.; Urushihara, M.; Takai, J.; Fujii, T.; Liang, X.G.; Kashida, H.; Asanuma, H.

    Chem. Sci.     2013年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: DOI:10.1039/C3SC51197A

  146. A "sugar-deficient" G-quadruplex: incorporation of aTNA in G4 structures 査読有り

    Zhou, J.; Murayama, K.; Amrane, S.; Kashida, H.; Bourdoncle, A.; Asanuma, H.; Mergny, J. L.

    Chem. Sci.   4 巻   頁: 3693-3698   2013年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  147. Highly stable duplex formation by artificial nucleic acids aTNA and SNA with acyclic scaffolds 査読有り

    Murayama, K.; Tanaka, Y.; Toda, T.; Kashida, H.; Asanuma, H.

    Chem. Eur. J.     2013年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/chem.201301578

  148. p-Stilbazole moieties as artificial base pairs for photocrosslinking of DNA duplex 査読有り

    Kashida, H.; Doi, T.; Sakakibara, T.; Hayashi, T.; Asanuma, H.

    J. Am. Chem. Soc.   135 巻   頁: 7960-7966   2013年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  149. Evaluation of intrinsic spectroscopic properties of chromophore assemblies by shielding with cyclohexyl base pairs within a DNA duplex 査読有り

    Kashida, H.; Higashiyama, N.; Kato, T.; Asanuma, H.

    Bioorg. Med. Chem.     2013年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.bmc.2013.04.032

  150. Photoswitch nucleic acid catalytic activity by regulating topological structure with a universal supra-photoswitch 査読有り

    Liang, X.G.; Zhou, M.G.; Kato, K.; Asanuma, H.

    ACS Synth. Biol   2 巻   頁: 194-202   2013年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  151. Development of a robust model system of FRET using base-surrogates tethering fluorophores for strict control of their position and orientation within DNA duplex 査読有り

    Kato, T.; Kashida, H.; Kishida, H.; Yada, H.; Okamoto, H.; Asanuma, H.

    J. Am. Chem. Soc.   135 巻   頁: 741-750   2013年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  152. Polycation-chaperoned in-stem molecular beacon system 査読有り

    Asanuma, H.; Osawa, T.; Kashida, H.; Fujii, T.; Liang, X. G.; Niwa, K.; Yoshida, Y.; Shimada, N.; Maruyama, A

    Chem. Commun.   48 巻   頁: 1760-1762   2012年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  153. Quencher-free linear probe with multiple fluorophores on acyclic scaffold 査読有り

    Asanuma, H.; Akahane, M.; Kondo, N.; Osawa, T.; Kato, T.; Kashida, H.

    Chem. Sci.     頁: in press   2012年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  154. Reversed assembling of dyes in RNA duplex compared with those in DNA 査読有り

    Fujii, T.; Urushihara, M.; Kashida, H.; Ito, H.; Liang, X.G.; Yagi-Utsumi, M.; Kato, K.; Asanuma, H.

    Chem. Eur. J.     頁: in press   2012年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  155. Bulge-like asymmetric hetero dye-clustering in DNA duplex results in efficient quenching of background emission based on the maximized excitonic interaction 査読有り

    Fujii, T.; Hara, Y.; Osawa, T.; Kashida, H.; Liang, X.G.; Yoshida, Y.; Asanuma, H.

    Chem. Eur. J.     頁: in press   2012年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  156. Quencher-free molecular beacon tethering 7-hydroxycoumarin detects targets through protonation/deprotonation 査読有り

    Kashida, H.; Yamaguchi K.; Hara, Y.; Asanuma, H.

    Bioorg. Med. Chem.   20 巻   頁: 4310-4315   2012年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  157. Coherent Interactions of Dyes Assembled on DNA 招待有り 査読有り

    Asanuma, H.; Fujii, T.; Kato, T.; Kashida, H.

    J. Photochem. Photobiol. C   13 巻   頁: 1065-1084   2012年

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    担当区分:筆頭著者   記述言語:英語  

  158. Preparation of supramolecular chromophoric assemblies using a DNA duplex 招待有り 査読有り

    Kashida, H.; Asanuma, H.

    Phys. Chem. Chem. Phys.   14 巻   頁: 7196-7204   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  159. Design of an Artificial Functional Nanomaterial with High Recognition Ability 招待有り 査読有り

    Liang, X. G.; Mochizuki, T.; Fujii, T.; Kashida, H.; Asanuma, H.

    Natural Computing   11 巻   頁: 231-238   2012年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  160. A model of elongation of short DNA sequence by thermophilic DNA polymerase under isothermal conditions 査読有り

    Kato, T.; Liang, X. G.; Asanuma, H.

    Biochemistry   51 巻   頁: 7846-7853   2012年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  161. Preparation of photoresponsive DNA tethering ortho-methylated azobenzene as a supraphotoswitch 招待有り 査読有り

    Asanuma, H.; Nishioka, H.; Ishikawa, T.; Liang, X. G.

    Current Protocols in Nucleic Acid Chemistry     頁: accepted   2011年9月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  162. Detection of three-base deletion by exciplex formation with perylene derivatives 査読有り

    Kashida, H.; Kondo, N.; Sekiguchi, K.; Asanuma, H.

    Chem. Commun.     頁: accepted   2011年7月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  163. Control of the Chirality and Helicity of Oligomers of Serinol Nucleic Acid (SNA) by Sequence Design 査読有り

    Kashida, H.; Murayama, K.; Toda, T.; Asanuma, H.

    Angew. Chem. Int. Ed.,   50 巻   頁: 1285-1288   2011年3月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  164. Cationic dye-triplet as unique "glue" that can connect full-matched termini of the duplexes 査読有り

    Kashida, H.; Hayashi, T.; Fujii, T.; Asanuma, H.

    Chem. Eur. J.   17 巻   頁: 2614-2622   2011年2月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  165. Improvement of RNAi activity and strand-selectivity of RISC formation by modified siRNA involving intercalators near 5'-termini 査読有り

    Hiroshi Ito, Masaaki Urushihara, Xingguo Liang, Hiroyuki Asanuma

    ChemBioChem     頁: ***-***   2011年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  166. Photon-fueled DNA nanodevice carrying two different photoswitches 査読有り

    Hidenori Nishioka, Xingguo Liang, Tomohiro Kato, Hiroyuki Asanuma

    Angew. Chem. Int. Ed.     頁: ***-***   2011年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  167. Cyclohexyl "base pairs" stabilize duplexes and intensify pyrene fluorescence by shielding it from natural base pairs 査読有り

    Kashida, H.; Sekiguchi, K.; Higashiyama, N.; Kato, T.; Asanuma, H.

    Org. Biomol. Chem.     頁: ***-***   2011年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  168. Ultrafast photoisomerization and its single-shot pump pulse efficiency of trans-azobenzene derivative: Compound for photosensitive DNA 査読有り

    T. Chen, A. Yamaguchi, K. Igarashi, N. Nakagawa, H. Nishioka, H. Asanuma, M. Yamashita

    Opt. Commun.     2011年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.optcom.2011.10.032

  169. Femtosecond photoisomerization of azobenzene-derivative binding to DNA 査読有り

    T. Chen, K. Igarashi, N. Nakagawa, K. Yamane, T. Fujii, H. Asanuma, M. Yamashita

    J. Photochem. Photobiol. A   223 巻   頁: 119-123   2011年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  170. Nick Sealing by T4 DNA Ligase on a Modified DNA Template Tethering a Functional Molecule on D-Threoninol 査読有り

    Liang Xingguo, Kenta Fujioka, Hiroyuki Asanuma

    Chem. Eur. J.   17 巻   頁: 10388-10396   2011年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  171. An interstrand-wedged duplex composed of alternating DNA base pairs and covalently attached intercalators 査読有り

    Liang, X.G.; Nishioka H.; Mochizuki, T.; Asanuma, H.

    J. Mater. Chem.   20 巻   頁: 575-581   2010年1月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    An interstrand-wedged duplex involving alternating base pairs and covalently attached intercalators via D-threoninols was constructed. In this novel duplex structure, natural DNA base pairs and artificially introduced planar molecules such as azobenzene derivatives are lined up one by one. Although each base pair is sandwiched by two intercalators and vice versa, the duplex is extremely stable compared with the corresponding native DNA duplex.

  172. An efficient FRET between pyrene and perylene assembled in a DNA duplex and its potential for discriminating single base changes 査読有り

    Kashida, H.; Takatsu,T.; Sekiguchi, K.; H. Asanuma

    Chem. Eur. J.   16 巻   頁: 2479-2486   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In order to increase the apparent Stokes' shift of perylene, pyrene (donor) and perylene (acceptor) were assembled in a DNA duplex to achieve the efficient fluorescence resonance energy transfer (FRET) from pyrene to perylene. Multiple donors were introduced in the vicinity of acceptors via D-threoninol and natural base pairs were inserted between the dyes. Accordingly, donors and acceptors could be accumulated inside the DNA without forming undesired excimer/exciplex. When two pyrene moieties were located in proximity to one perylene with one base pair inserted between them, efficient FRET occurred within the duplex. Thus, strong emission at 460 nm was observed from perylene when excited at 345 nm where pyrene has its absorption. The apparent Stokes' shift became as large as 115 nm with a high apparent FRET efficiency.

  173. Construction of photoresponsive RNA for photoswitching RNA hybridization 査読有り

    Ito, H.; Liang, X.G.; Nishioka, H.; Asanuma, H.

    Org. Biomol. Chem.     頁: in press   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  174. Robust and photo-controllable DNA capsules using azobenzenes 査読有り

    Tanaka, F.; Mochizuki, T.; Liang, X.G.; Asanuma, H.; Tanaka, S.; Suzuki, K.; Kitamura, S.; Nishikawa, A.; Ui-Tei, K.; Hagiya, M

    Nano Lett.   10 巻   頁: 3560-3565   2010年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  175. Unexpectedly stable artificial duplex from flexible acyclic threoninol. 査読有り

    Asanuma, H.; Toda, T.; Murayama, K.; Liang, X.G.; Kashida, H.

    J. Am. Chem. Soc.   132 巻   頁: 14702-14703   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  176. Insulator base pairs for lighting-up perylenediimide in a DNA duplex 査読有り

    Hiromu Kashida,Koji Sekiguchi,and Hiroyuki Asanuma

    Chem. Eur. J.   16 巻   頁: 11554-11557   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Perylenediimide (PDI) is highly quenched by nucleobases, which greatly restricts its application as a fluorescent probe. Here, we propose “insulator base pairs" tethering cyclohexane ring through D-threoninol. When “insulator base pairs" were inserted between PDI and nucleobases, the quantum yield of PDI drastically increased several thousand-fold. The “insulator base pairs" reported here also have the potential to increase the quantum yields of other fluorophores.

  177. Coherent Quenching of a Fluorophore for the Design of a Highly Sensitive In-Stem Molecular Beacon. 査読有り

    Yuichi Hara, Taiga Fujii, Hiromu Kashida, Koji Sekiguchi, Xingguo Liang, Kosuke Niwa, Tomokazu Takase, Yasuko Yoshida, and Hiroyuki Asanuma

    Angew. Chem. Int. Ed.   49 巻   頁: 5502-5506   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Excitonic interaction (coherency) was utilized to design a highly sensitive In-Stem molecular beacon (ISMB) in which both a fluorophore and a quencher on D-threoninols are incorporated into the stem region as a pseudo “base-pair". A systematic study indicated that minimization of the difference of &#61548;max between the fluorophore and the quencher maximized quenching efficiency due to maximization of coherency. A highly sensitive ISMB could be prepared using the excitonically optimized pair of Cy3 and modified Methyl Red.

  178. Accumulation of fluorophores into DNA duplexes to mimic the properties of quantum dots 査読有り

    Hiromu Kashida, Koji Sekiguchi, Xingguo Liang, Hiroyuki Asanuma

    J. Am. Chem. Soc.   132 巻   頁: 6223-6230   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    By using perylene and pyrene as fluorophores, we have designed various fluorophore assemblies that mimic inorganic quantum dots in showing a high emission intensity, a large Stokes' shift, and a modulated emission maximum. For this purpose, we utilized two kinds of duplex motifs with D-threoninols as scaffolds: cluster and interstrand-wedged motifs. In the cluster motif, fluorophores are introduced into both strands to produce tentative pseudo-“base-pairs", in which the dyes strongly interact with each other and form dimers, trimers or hexamers. In the interstrand-wedged motif, a base-pair is inserted between the fluorophores to suppress their direct interaction. These two motifs were applied to accumulate dyes within a DNA duplex depending on their emission properties. Since pyrene exhibits strong excimer emission, the emission at 500 nm of a pyrene cluster motif strongly increased as the number of accumulated dyes increased, whereas interstrand-wedged motif quenched pyrene monomer emission. In contrast, assembled perylenes, which are mostly quenched by dimerization, showed intense monomer emission in the interstrand-wedged motif whereas perylene cluster motifs strongly suppressed perylene emission. These two motifs were then applied to the hetero-assembly of pyrenes and perylenes. Both a large Stokes' shift and a modulation of the emission maximum, which are also characteristics of inorganic quantum dots, were successfully realized using fluorescent resonance energy transfer (FRET) and exciplex formation. These fluorophore assemblies thus obtained could be enzymatically ligated to longer DNA, demonstrating that this technique has the potential to be a versatile labeling agent for biomolecules.

  179. *Photoregulation of DNA transcription by using photoresponsive T7 promoter and clarification of its mechanism 査読有り

    Xingguo Lianga, Ryuji Wakuta, Kenta Fujioka, Hiroyuki Asanuma

    FEBS Journal   277 巻   頁: 1551-1561   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Using photoresponsive T7 promoters tethering two 2&#61602;-methylazobenzenes or 2&#61602;,6&#61602;-dimethylazobenzenes, highly efficient photoregulation of DNA transcription was attained. Under UV-A irradiation (320-400 nm), the rate of transcription with T7 RNA polymerase and a photoresponsive promoter involving two 2&#61602;,6&#61602;-dimethylazobenzenes was 10-fold faster than that after visible light irradiation (400-600 nm). By attaching a non-modified azobenzene (Azo) and 2&#61602;,6&#61602;-dimethylazobenzene (DM-azo) at the two positions, respectively, and by utilizing the different cis-to-trans thermal stability between cis Azo and cis DM-azo, four species of T7 promoter (cis-cis, trans-cis, cis-trans, and trans-trans) were obtained. The four species showed transcriptional activity in the order of cis-cis > cis-trans > trans-cis > trans-trans. Kinetic analysis revealed that the Km for the cis-cis promoter (both of the introduced azobenzene derivatives were in the cis form) and T7 RNA polymerase was 68 times larger than that of the trans-trans form, indicating that high photoregulatory efficiency was mainly due to a remarkable difference in affinity with RNA polymerase. The present approach is promising for the creation of biological tools for artificially controlling gene expression, and as a photo-controlled system for supplying RNA fuel for RNA-powered molecular nanomachines.

  180. *A light-driven DNA nanomachine for efficiently photoswitching RNA digestion 査読有り

    Zhou M.G.; Liang, X.G.; Mochizuki, T.; Asanuma, H.

    Angew. Chem. Int. Ed.   49 巻   頁: 2167-2170   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A machine-like photoresponsive DNA enzyme was constructed that can work for us at a single molecule level. The complete ON-OFF photoswitching of RNA digestion was realized by photoregulating the topological structure of DNAzyme/RNA complex. The interstrand-wedged duplex was used as the supra-photoswitch.

  181. Effect of the ortho modification of azobenzene on the photoregulatory efficiency of DNA hybridization and thermal stability of its cis-form 査読有り

    Nishioka H.; Liang, X.G.; Asanuma, H.

    Chem. Eur. J.   16 巻   頁: 2054-2062   2010年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We synthesized various azobenzenes methylated at their ortho-positions with respect to the azo bond for more effective photoregulation of DNA hybridization. Photoregulatory efficiency, evaluated from the change of Tm (&#61508;Tm) induced by trans-cis isom-erization, was significantly improved for all ortho-modified azobenzenes compared with non-modified azoben-zene due to the more stabilized trans-form and more destabilized cis-form. Among the synthesized azobenzenes, 4-carboxy-2&#61602;,6&#61602;-dimethylazobenzene (2&#61602;,6&#61602;-Me-Azo), in which two ortho-positions of the distal benzene ring with respect to carboxyl group were methy-lated, exhibited the largest &#61508;Tm, whereas newly synthesized 2,6-Me-Azo (4-carboxy-2,6-dimethylazobenzene), which possesses two methyl groups on the two ortho-positions of the other benzene ring, showed moderate im-provement of &#61508;Tm.

  182. Line up base pairs and intercalators one by one in a stable duplex

    Liang, X.G.; Mochizuki T.; Nishioka H.; Asanuma, H.

    Nucleic Acids Symp. Ser.   53 巻   頁: 189-190   2009年11月

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    担当区分:筆頭著者   記述言語:英語  

    A stable double helix involving alternating base pairs and azobenzene moieties was constructed. In this supramolecule, base pairs and azobenzenes are lined up one by one to form an interstrand-wedged motif: each base pair is sandwiched with two azobenzenes, and each azobenzene intercalates between two base pairs. This motif was formed by the hybridization of two modified DNA tethering multiple azobenzene moieties at a frequency of one azobenzene for every two nucleotides. Furthermore, this structure could be simply dismantled by UV light irradiation and reformed with the irradiation of visible light. By using this new duplex motif, construction of a variety of photoresponsive nanostructures and nanodevices is highly expected.

  183. Efficient energy transfer from pyrene to perylene assembled inside DNA duplex

    Kashida, H.; Takatsu, T.; Asanuma, H.

    Nucleic Acids Symp. Ser.,   53 巻   頁: 29-30   2009年11月

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    担当区分:筆頭著者   記述言語:英語  

    Pyrene (donor) and perylene (acceptor) were assembled in a DNA duplex to increase the apparent Stokes' shift of perylene. Plural Multiple donors were introduced in the vicinity of acceptors via D-threoninol and natural base pairs were inserted between donors and acceptors in order to suppress undesired interactions between them. When two pyrene moieties were located in proximity to one perylene with one base pair inserted between them, efficient FRET occurred within the duplex. Thus, strong emission at 495 nm was observed from perylene when excited at 345 nm where pyrene has its absorption. The apparent Stokes' shift became as large as 115 nm with a high FRET efficiency (&#61510;>1). However, the introduction of more than two pyrenes did not enhance the fluorescence intensity of perylene, due to the short F&ouml;rster radius (R0) of the donor pyrene.

  184. *Analysis of Coherent Heteroclustering of Different Dyes by Use of Threoninol-Nucleotides for Comparison with the Molecular Exciton Theory 査読有り

    Taiga Fujii, Hiromu Kashida, Hiroyuki Asanuma

    Chem. Eur. J.   15 巻   頁: 10092-10102   2009年10月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    To test the molecular exciton theory for heterodimeric chromophores, various heterodimers and clusters, in which two different dyes were stacked alternately, were prepared by hybridizing two oligodeoxyribo-nucleotides (ODNs), each of which tethered a different dye on D-threoninol at the center of the strand. NMR analyses revealed that two different dyes from each strand were stacked antiparallel to each other in the duplex, and were located adjacent to the 5'-side of a natural nucleobase. Spectroscopic behaviors of these heterodimers were systematically examined as a function of the difference in the wavelength of the dye absorption maxima (&#61508;&#61548;max). We found that the absorption spectrum of the heterodimer was significantly different from that of the simple sum of each monomeric dye in the single-strand. When azobenzene and Methyl Red, which have &#61548;max at 336 nm and 480 nm in the single-strand, respectively, (&#61508;&#61548;max = 144 nm), were assembled on ODNs, the band derived from azobenzene exhibited a small hyperchromism whereas the band from Methyl Red showed hypochromism and both bands shifted to a longer wavelength (bathochromism). These hyper- and hypochromisms were further enhanced in a heterodimer derived from 4'-methylthioazobenzene and Methyl Red that had a much smaller &#61508;&#61548;max (82 nm) (&#61548;max = 398 and 480 nm in the single-strand, respectively). With a combination of 4'-dimethylamino-2-nitroazobenzene and Methyl Red, which had an even smaller &#61508;&#61548;max (33 nm), a single sharp absorption band that was apparently different from the sum of the single-stranded spectra was observed. These changes in the intensity of the absorption band could be explained by the molecular exciton theory that has been mainly applied to the spectral behavior of H- and/or J-aggregates composed of homo dyes. However, the bathochromic band shifts observed at shorter wavelengths did not agree with the hypsochromism predicted by the

  185. *In-Stem Molecular Beacon containing a Pseudo Base Pair of Threoninol Nucleotides for Removal of Background Emission 査読有り

    Hiromu Kashida, Tomohiko Takatsu, Taiga Fujii, Koji Sekiguchi, Xingguo Liang, Kosuke Niwa, Tomokazu Takase, Yasuko Yoshida, and Hiroyuki Asanuma

    Angew. Chem. Int. Ed.   48 巻   頁: 7044-7047   2009年9月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    An in-stem molecular beacon (IS-MB), which tethers perylene and anthraquinone in the stem region via D-threoninol, effectively detected target sequences. This design of IS-MB is also applicable to the discrimination of a one-base deletion mutant from wild type (full-match) without background emission.

  186. Photoisomerization dynamics study on cis-azobenzene derivative using ultraviolet-to-visible tunable femtosecond pulses 査読有り

    Yamaguchi, N.; Nagasawa, N.; Igarashi, K.; Sekikawa, T.; Asanuma, H.; Yamashita, M.

    Appl. Surf. Sci.   255 巻   頁: 9864-9868   2009年7月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We performed the transient absorption measurement and the first rate equation (RE) analysis for cis isomer of 4-carboxy-20,60-dimethylazobenzene to clarify the quantitative difference between the photoisomerization process and the thermal relaxation process from the SC
    1 excited state. The RE analysis enabled us to determine the cis-to-trans photoisomerization rate per each pump pulse to be 3% under the condition of the 430 nm, 150 fs pump pulse with energy of 200 nJ. Moreover, the signal due to the yielded trans molecules appearing in the transient absorption was assigned from the following observed result: the transient absorbance change at the 380 nm probe mostly decreased within 300 fs after the 430 nm pulse pumping and then slowly decreased to zero, while the absorbance change at the 350 nm probe had a positive constant component in the over one picosecond time region. The RE analysis showed
    that this constant component is due to the yielded transmolecules, and its positive value is due to the fact
    that the absorption cross-section of the ST 0 -to- ST
    2 transition in their trans molecules is larger than that of
    the SC0 -to- SC 2 transition in the original cis molecules.

  187. Positively charged base surrogate for highly stable “base-pairing" through electrostatic and stacking interactions. 査読有り

    Hiromu Kashida, Hidehiro Ito, Taiga Fujii, Takamitsu Hayashi, Hiroyuki Asanuma

    J. Am. Chem. Soc.   131 巻   頁: 9928-9930   2009年7月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    “Base pairs" of cationic dyes (p-methylstilbazole) were incorporated into oligodeoxyribonucleotides (ODNs). This “base pair" greatly stabilized the duplex through electrostatic and stacking interactions. The melting temperature of modified ODN was higher than those of neutral dyes and native base pairs. Further stabilization of the duplex was observed when the number of cationic dyes increased.

  188. *A Supra-Photoswitch Involving Sandwiched DNA Base Pairs and Azobenzenes for Light-Driven Nanostructures and Nanodevices 査読有り

    Liang, X.G.; Mochizuki, T.; Asanuma, H.

    Small   5 巻 ( 15 ) 頁: 1761-1768   2009年6月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A supra-photoswitch was designed for complete ON / OFF switching of DNA hybridization by light irradiation for the purpose of using DNA as a material for building nanostructures. Azobenzenes, attached to D-threoninols that function as scaffolds, were introduced into each DNA strand after every two natural nucleotides (in the form (NNX)n where N and X represent the natural nucleotide and the azobenzene moiety, respectively). Hybridization of these two modified strands formed a supra-photoswitch consisting of alternating natural base pairs and azobenzene moieties. In this newly designed sequence, each base pair is sandwiched between two azobenzene moieties and all the azobenzene moieties are separated by base pairs. When the duplex is irradiated by visible light, the azobenzene moieties take the trans form and this duplex is surprisingly stable compared to the corresponding native duplex composed of only natural oligonucleotides. On the other hand, when the azobenzene moieties are isomerized to the cis form by UV light irradiation, the duplex is completely dissociated. Based on this design, a DNA hairpin structure was synthesized that should be closed by visible light irradiation and opened by UV light irradiation at the level of a single molecule. Indeed, perfect ON-OFF photoregulation was attained. This design is a promising strategy for the design of supra-photoswitches such as photoresponsive sticky ends on DNA nano devices and other nanostructures.

  189. Construction of Photon-Fueled DNA Nanomachines by Tethering Azobenzenes as Engines 査読有り

    Liang, X.G.; Nishioka, H.; Takenaka, N.; Asanuma, H.

    Lecture Note in Computer Science   5347 巻   頁: 21-32   2009年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Nanoscale DNA tweezers operated by photo-irradiation were constructed by using azobenzene-modified DNA as materials. The azobenzenes that can photoisomerize between trans and cis form were used as the engines to open and close the tweezers. The work principle is based on the reversible photoregulation of DNA hybridization. When non-substituted azobenzene was used, the tweezers were open after UV light irradiation ((330-350 nm, cis form), and closed after visible light irradiation (440-460 nm, trans form). More interestingly, the operation reversed when an azobenzene derivative with a para-isopropyl group was used: UV light irradiation closed the tweezers and visible light irradiation opened them. As compared with the oligonucleotide-fuelled DNA machines, the nanomachines constructed here were “environment-friendly" because no dsDNA waste was produced. Furthermore, the operation can be repeated many times simply by switching the photoirradiation without any decrease of the cycling efficiency.

  190. DNAを切断するはさみ

    西岡英則、浅沼浩之

    化学   64 巻   頁: 70-71   2009年

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    担当区分:筆頭著者   記述言語:日本語  

    遺伝子工学(遺伝子組み換え技術)は、1)遺伝子であるDNAの特定の位置での“裁断”、2)得られたDNA断片の、ベクターと呼ばれる遺伝子の“運び屋”への“縫製”、そして3)ベクターの細胞内への導入(形質転換)、という3つの基本技術に基づいている。この中でも1)のDNAを“裁断”する “はさみ”=制限酵素 の発見が、現在の遺伝子工学を可能にしたと言っても過言ではない。この天然由来の“はさみ”に相当する制限酵素には様々な種類が存在し、その多くは4~6塩基程度の塩基配列を認識してDNAを切断する。したがって数千塩基程度の短い遺伝子ならば、天然の制限酵素を用いることで特定の1~2箇所を切断することが可能である。しかしながら、高等生物のような巨大なゲノムDNAに対して利用すると非常に多くの断片が生じてしまう。例えば、30億塩基を持つヒトゲノムに対して6塩基しか認識しない制限酵素を利用すると、おおよそ73万 (= 30億÷46)もの断片が生じてしまうことになる。すなわち、天然の制限酵素では、高等生物のDNAを配列特異的に1箇所で切断することは不可能である。このような問題点を克服するために近年、認識配列に制限が無い人工的な “はさみ”=「人工制限酵素」に関する様々な研究が活発に行われている。ここでは、DNAを配列特異的に切断する人工制限酵素について紹介する。

  191. *Rational Design of Functional DNA with a Non-Ribose Acyclic Scaffold 招待有り 査読有り

    H. Kashida, X.G. Liang, H. Asanuma

    Current Organic Chemistry   13 巻 ( 11 ) 頁: 1065-1084   2009年

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    担当区分:筆頭著者   記述言語:英語  

    The growing field of DNA technology requires new modified DNAs that can perform advanced functions. No matter how we optimize the length and sequence of DNA using only the four naturally occurring nucleotides, potential performance is limited. In this review, we describe a facile and effective method of rationally designing new functional DNA by focusing on acyclic scaffolds, especially threoninols, which are utilized to incorporate functional molecules into
    DNA. Wedge-type insertion of a functional molecule with a planar structure of proper size in D-threoninol to DNA does
    not destabilize the duplex, although the backbone structure is changed. Rather, intercalation offsets such distortions and significantly raises the melting temperature of the DNA duplex. Based on the wedge-type insertion, photoresponsive DNA (tethering azobenzenes) and fluorescent probes that can detect single nucleotide polymorphisms (SNPs) and insertion/
    deletion (indel) polymorphisms have been designed. Furthermore, a variety of molecular clusters of dyes have also been prepared from acyclic scaffolds tethering dyes.

  192. Glucuronidase-assisted transglycosylation for the synthesis of highly functional disaccharides: β-D-Glucuronyl 6-O-sulfo-β-D-gluco- and -β-D-Galactopyranosides 査読有り

    Nagatsuka T.; Uzawa H.; Asanuma H., Nishida, Y.

    J. Carbohydrate Chem.   28 巻   頁: 94-106   2009年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The substrate specificity of snail, limpet, and bovine glucuronidases was examined by using p-nitrophenyl glucuronide and p-nitrophenyl 6-O-sulfo-D-glycopyranosides as the glycosyl donor and acceptors, respectively. When the donor was treated with these enzymes in the absence of the acceptors, beta(1-3) glucuronyl disaccharides were obtained as the major products together with beta(1-2) isomers as the result of an enzymatic "self-transglycosylation"reaction.

  193. Modulation of pKa of Brooker's Merocyanine by DNA Hybridization 査読有り

    Kashida, H.; Sano, K.; Hara, Y.; Asanuma, H.

    Bioconjugate Chem.   20 巻   頁: 258-265   2009年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Brooker's merocyanine (BM), which changes its emission and absorption maxima upon protonation, was introduced into oligodeoxyribonucleotide (ODN) via D-threoninol by postsynthetic modification on a CPG (Controlled-Pore Glass) support. The pKa of BM in the modified ODN increased from 9.5 to 10.1 upon hybridization. As a result, absorption maxima shifted from 492 nm to 432 nm at pH 10.0 by the presence of its complementary strand. This spectral shift was sufficiently large so that DNA hybridization could easily be discriminated even by the naked eye; the color of the solution changed from orange to yellow upon hybridization. In addition, the fluorescence emission was strongly quenched upon hybridization, demonstrating that this probe can also detect the target DNA by the fluorescence change. Ratiometric detection of hybridization was also possible by simultaneous excitation of both protonated and deprotonated BMs. Furthermore, we could also modulate its pKa by the anti-parallel stacking of two BM molecules in the duplex; the pKa of BM decreased from 10.1 to 9.7 by the stacking of two BMs in an anti-parallel manner. Thus, control of the microenvironment around the BM molecule allowed modulation of its pKa, which is applicable to the sequence specific recognition of target DNA.

  194. Light driven open/close operation of an azobenzene-modified DNA nano-pincette

    Liang, X.G.; Takenaka, N.; Nishioka, H.; Asanuma, H.

    Nucleic Acids Res. Symp. Ser.   52 巻   頁: 697-698   2008年9月

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    担当区分:筆頭著者   記述言語:英語  

    A photoresponsive DNA nano-pincette was constructed by using azobenzene-modified DNA as materials. When the azobenzene-modified part hybridizes with its complementary sequence on the pincette, the duplex formation closes it. On the contrary, the pincette is opened after the formed duplex dissociates. Based on reversible photoswitching of this DNA hybridization, the pincette involving non-substituted azobenzene can be opened simply by UV light irradiation and closed by visible light irradiation. Interestingly, the operation can be reversed by using para-isopropyl group substituted azobenzene: visible light opens the pincette, and UV light closes it. In both cases, the azobenzene-modified part was attached to the pincette throughout the open/close operation, which makes single molecular operation possible. Furthermore, the operation can be repeated many times without any decrease of the cycling efficiency and no DNA waste was produced.

  195. Preparation of coherent hetero clusters with threoninol scaffold.

    Fujii, T.; Kashida, H.; Asanuma, H.

    Nucleic Acids Symp. Ser.   52 巻   頁: 699-700   2008年9月

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    担当区分:筆頭著者   記述言語:英語  

    New hetero aggregates where different dyes stacked alternately were prepared by hybridizing two DNAs, each of which tethered a different dye in the centre of strand. Spectral changes due to exciton coupling between different kinds of dyes were observed. Especially, hetero aggregates of Methyl Red and 2-nitro-4'-dimethylaminoazobenzene showed substantial narrowing of the band, demonstrating coherent coupling occurred in these aggregates.

  196. Incorporation of cationic dyes into DNA for distinct stabilization of duplex

    Kashida, H.; Itoh, H.; Fujii, T.; Asanuma, H.

    Nucleic Acids Symp. Ser.   2008 巻   頁: 701-702   2008年9月

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    担当区分:筆頭著者   記述言語:英語  

    In this study, cationic dyes (methylstilbazole) were introduced into ODN. When two complementary ODNs, both of which tethered thise dye, were hybridized, the melting temperature drastically increased. Furthermore, The duplex was further stabilized by introducing multiple dyes.

  197. Construction of a photo-switchable gene for turning on and off gene expression with light irradiation

    Liang, X.G.; Fujioka, K.; Tsuda, Y.; Wakuda, R.; Asanuma, H.

    Nucleic Acids Res. Symp. Ser.   52 巻   頁: 19-20   2008年9月

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    担当区分:筆頭著者   記述言語:英語  

    A photoresponsive GFP gene was constructed by attaching a T7 promoter that involves two azobenzene moieties as the photoswitch. The azobenzene moieties tethered on D-threoninol were inserted precisely into the sequence of T7 promoter at two positions in the non-template strand. By using azobenzene-tethered DNA as a primer, azobenzene was attached to GFP gene after PCR amplification. However, a single-stranded overhang involving azobenzene was formed because primer extension stopped at the position of azobenzene moiety. Interestingly we found that oligonucleotide complementary to the overhang could be ligated by T4 DNA ligase at the stopped position, and the intact photoresponsive T7 promoter was attached onto GFP gene. Furthermore, the in vitro expression of the constructed photoresponsive GFP gene was successfully switched on and off with light irradiation.

  198. A DNA Nanomachine Powered by Light Irradiation 査読有り

    Liang, X.G.; Takenaka, N.; Nishioka, H.; Asanuma, H.

    ChemBioChem   9 巻 ( 5 ) 頁: 702-705   2008年5月

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The photoresponsive DNA tweezers as a nanomachine fuelled with photons were constructed by using azobenzene-modified DNA. The tweezers were photoswitched to be open with the UV light irradiation (335-345 nm) and to be closed with the visible light irradiation (445-455 nm) without further adding oligonucleotides as the fuel.

  199. Postsynthetic modification of DNA via threoninol on a solid support by means of allylic protection 査読有り

    Hiroyuki Asanuma, Yuichi Hara, Akira Noguchi, Kanae Sano, Hiromu Kashida

    Tetrahedron Letters   49 巻   頁: 5144-5146   2008年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We have developed a facile but versatile method to introduce functional molecules into DNA on a CPG support. Threoninol, whose amino group was protected with an (allyloxy)carbonyl (Alloc) group, was introduced into DNA via the corresponding phosphoramidite monomer. After selective deprotection of the Alloc group by treatment with palladium(0), a dye with a carboxyl group could be introduced into the DNA on the CPG support through an amide bond.

  200. Threoninol as a Scaffold of Dyes (Threoninol-nucleotide) and Their Stable Interstrand Clustering in Duplexes 査読有り

    Hiromu Kashida, Taiga Fujii, Hiroyuki Asanuma

    Org. Biomol. Chem.   6 巻 ( 16 ) 頁: 2892 - 2899   2008年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Functional molecules such as dyes (Methyl Red, azobenzene, and Naphthyl Red) were tethered on D-threoninol as base surrogates (threoninol-nucleotide), which were consecutively incorporated at the center of natural oligodeoxyribonucleotides (ODNs). Hybridization of two ODNs involving threoninol-nucleotides allowed interstrand clustering of the dyes on D-threoninol and greatly stabilized the duplex. When two complementary ODNs, both of which had tethered Methyl Reds on consecutive D-threoninols, were hybridized, the melting temperature increased proportionally to the number of Methyl Reds, due to stacking interactions. Clustering of Methyl Reds induced both hypsochromicity and narrowing of the band, demonstrating that Methyl Reds were axially stacked relative to each other (H-aggregation). Since hybridization lowered the intensity of circular dichroism peaks at the p-p* transition region of Methyl Red (300 nm – 500 nm), clustered Methyl Reds were scarcely wound in the duplex. Alternate hetero dye clusters could also be prepared only by hybridization of two ODNs with different threoninol-nucleotides, such as Methyl Red / azobenzene and Methyl Red / Naphthyl Red combinations. A combination of Methyl Red and azobenzene induced bathochromic shift and broadening of the band at the Methyl Red region due to the disturbance of exciton interaction among Methyl Reds. But interestingly, the Methyl Red and Naphthyl Red combination induced merging of each absorption band to give a single sharp band, indicating that exciton interaction occurred among the different dyes. Thus, D-threoninol can be a versatile scaffold for introducing functional molecules into DNA for their ordered clustering.

  201. Molecular Design for Reversing the Photoswitching Mode of Turning ON and OFF DNA Hybridization 査読有り

    Xingguo Liang, Nobutaka Takenaka, Hidenori Nishioka, Hiroyuki Asanuma

    Chem. Asian J.   9 巻   頁: 702-705   2008年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A new photoswitch of DNA hybridization involving para-substituted azobenzene (such as isopropyl or tert-butyl substituted one) on L-threoninol as a linker was synthesized. When visible light was irradiated to the modified DNA, the duplex was dissociated due to the destabilization effect of the bulky substituent on trans-azobenzene. In contrast, trans-to-cis isomerization (UV light irradiation) facilitated the duplex formation. The direction of this photoswitching mode was entirely reversed as compared with the previous one carrying an unmodified azobenzene on D-threoninol whose trans-form turned on the hybridization, and cis-form turned it off. Such reversed and reversible photoswitching of DNA hybridization was directly demonstrated by using fluorophore- and quencher-attached oligonucleotides. Furthermore, it was revealed that the cis-to-trans thermal isomerization was greatly suppressed in the presence of the complementary strand due to the formation of more stable duplex in cis-form.

  202. Diastereomer Separation of Azobenzene-Tethered Oligodeoxyribonucleotides and Determination of Their Absolute Configurations by Enzymatic Digestion. 査読有り

    Xingguo Liang, Makoto Komiyama, Hiroyuki Asanuma

    Nucleosides, Nucleotides & Nucleic Acids   27 巻   頁: 332-350   2008年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Two diastereomers were produced by the introduction of azobenzene-tethering prochiral linker (2,2-bis(hydroxymethyl)propionic acid) in the modified ODN, which had been used for the photo-regulation of DNA functions. We found that this modified ODN with sequence 5&cent;-…pNpXpN…-3&cent; (p = phosphate; N = nucleoside; X = azobenzene residue) could be digested to pX (the phosphate at the 5&cent; side of X was left) by an over excess of Phosphodiesterase I. By comparing the retention time of pX from the separated diastereomer with that of authentic R- or S-pX on chiral HPLC, absolute configuration could be easily determined.

  203. DNA-色素コンジュゲーションによるくし型へテロ会合体の調製

    藤井大雅、樫田啓、浅沼浩之

      51 巻   頁: 277-278   2007年11月

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    担当区分:筆頭著者   記述言語:英語  

  204. Photoregulation of DNA hybridization by introducing an azobenzene: Molecular design for more stabilization of DNA duplex with cis-azobenzene than with its trans-form.

    Xingguo Liang, Nobutaka Takenaka, Hidenori Nishioka, Hiroyuki Asanuma

    Nucleic Acids Symp. Ser.   51 巻   頁: 169-170   2007年11月

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    担当区分:筆頭著者   記述言語:英語  

    Previously, photoregulation of DNA hybridization was achieved by introducing nonsubstituted azobenzene via a D-threoninol linker: DNA duplex formed (ON) after visible light irradiation (planar trans-form), whereas the duplex dissociated (OFF) after UV light irradiation (non-planar cis-form). In this study, for more efficient photoregulation of DNA functions, the reverse switch that can turn on duplex formation with UV, and turn off it with visible light irradiation was designed. When para-isopropylazobenzene (p-iPrAzo) was introduced into DNA via a L-thereoninol linker, the photoswitching direction was completely reversed: the duplex involving non-planar cis-p-iPrAzo was much more stable than that involving planar trans-form.

  205. Unexpected efficient ab initio DNA synthesis at low temperature by using thermophilic DNA

    Xingguo Liang, Tomohiro Kato, Hiroyuki Asanuma

    Nucleic Acids Symp. Ser.   51 巻   頁: 351-352   2007年11月

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    担当区分:筆頭著者   記述言語:英語  

    DNA was synthesized in the absence of DNA or RNA as template (or primer) from dNTPs at relatively low temperatures (25~50oC) by thermophilic Vent DNA polymerase whose proper reaction temperature for primer extension is 70~80oC. Unexpectedly, the ab initio DNA synthesis was even more efficient at 50oC as compared with that at 70oC. Interestingly, the ab initio DNA synthesis by Vent (exo-), a mutant version of Vent DNA polymerase lacking of 3&cent;&reg;5&cent; exonuclease activity, became much less efficient, and it could only carry out ab intio DNA synthesis after a long incubation time. This remarkable difference between Vent and Vent (exo-) indicates that the exonuclease activity domain of Vent may play an important role at the initiation step of ab initio DNA synthesis.

  206. Effective photoregulation of gene expression by photoresponsive T7 promoter

    Xingguo Liang, Ryuji Wakuda, Yuichiro Tsuda, Hidenori Nishioka, Hiroyuki Asanuma

    Nucleic Acids Symp. Ser.   51 巻   頁: 349-350   2007年11月

     詳細を見る

    担当区分:筆頭著者   記述言語:英語  

    We constructed a photoresponsive T7 promoter by tethering two 2,6-dimethyl azobenzenes (4-(2,6-dimethyl-phenlylazo)-benzoic acids) via D-threoninol linkers. Under UV light irradiation, the rate of transcription with T7 RNA polymerase (RNAP) on the photoresponsive promoter was 10-fold faster than that under visible light irradiation. Kinetic analysis revealed that Km of cis-cis-promoter (both of the introduced azobenzenes are in cis-form) with T7 RNAP was more than 60 times larger than that of trans-trans-form, indicating that the high photoregulatoryion efficiency was obtained mainly due to the remarkable difference in their affinity with RNAP. By attaching a photoresponsive promoter to the GFP gene, we also showed that photoregulation of gene expression became practicable.

  207. Development of photoresponsive RNA towards photoswitching of RNA functions

    Hiroshi Ito, Hidenori Nishioka, Xingguo Liang, Hiroyuki Asanuma

    Nucleic Acids Symp. Ser.   51 巻   頁: 171-172   2007年11月

     詳細を見る

    担当区分:筆頭著者   記述言語:英語  

    By introducing azobenzene into RNA via a D-threoninol linker, the photoresponsive RNA was developed for the photoswitching of RNA hybridization. The Tm measurements showed that RNA/RNA duplex formation and dissociation could be efficiently photoregulated. The difference in Tm between trans- and cis-form (DTm) was as large as 9~12oC when azobenzene was tethered at the central position of a 10-nt-long RNA. Interestingly, photoregulation ability of the azobenzene introduced in RNA was even greater than that in DNA for photoswitching the corresponding duplex formation. The constructed photoresponsive RNA is promising to be applied for the photoregulation of RNA functions such as Ribozyme activity, RNAi, pre-mRNA editing, and aptamer complex formation.

  208. Design of peptide nucleic acid by introduction of carbohydrate on lysine linker.

    Akira Noguchi, Hiroyuki Asanuma

      51 巻   頁: 261-262   2007年11月

     詳細を見る

    担当区分:筆頭著者   記述言語:英語  

    We synthesized “cartridge" type monomer involving mannose unit on lysine as a linker and additionally in-serted it to peptide nucleic acid (PNA) oligomer. Ad-vantage of this procedure is that functional molecule could be easily introduced into anywhere position of PNA without sacrificing base-pairs, although such in-sertion lowered stability of the duplex with DNA.

  209. ペリレンを使用した高感度DNAプローブの開発

    樫田啓、髙津智彦、浅沼浩之

      51 巻   頁: 279-280   2007年11月

     詳細を見る

    担当区分:筆頭著者   記述言語:英語  

  210. Detection of genetic polymorphisms with high sensitivity by DNA-perylene conjugate 査読有り

    Hiromu Kashida, Tomohiko Takatsu, Hiroyuki Asanuma

    Tetrahedron Letters   48 巻   頁: 6759-6762   2007年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Modified oligodeoxyribonucleotides (ODNs) involving two perylene moieties are synthesized. By using this ODN, one-base deletion can easily be distinguished with high sensitivity. In addition, emission color of the solution greatly changed so that the detection was possible even by naked eyes.

  211. 2&cent;,6&cent;-Dimethylazobenzene as an efficient and thermo-stable photo-regulator for the photoregulation of DNA hybridization 査読有り

    Hidenori Nishioka, Xingguo Liang, Hiromu Kashida, and Hiroyuki Asanuma

    Chem. Commun.   2007 巻   頁: 4354-4356   2007年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The introduction of methyl groups into two ortho positions (2&cent; and 6&cent; positions) of the same benzene ring in an azobenzene remarkably raised both its photoregulation ability and the thermal stability of the cis-form.

  212. Recognition of Solution Structures of Peptides byMolecularly Imprinted Cyclodextrin Polymers 査読有り

    Shi-hui Song, Kazumi Shirasaka, Mami Katayama,Suguru Nagaoka, Shinji Yoshihara, Tomo Osawa,Jun Sumaoka, Hiroyuki Asanuma, andMakoto Komiyama

    Macromolecules   40 巻   頁: 3530-3532   2007年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  213. Synthesis of azobenzene-tethered DNA for reversible photo-regulation of DNA functions: hybridization and transcription 招待有り 査読有り

    Hiroyuki Asanuma, Xingguo Liang, Hidenori Nishioka, Daijiro Matsunaga, Minghe Liu, Makoto Komiyama

    Nature Protocols   2 巻 ( 1 ) 頁: 203-212   2007年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A phosphoramidite monomer bearing an azobenzene is synthesized from D-threoninol. By using this monomer, azobenzene moieties can be introduced into oligodeoxyribonucleotide (DNA) at any position on a conventional DNA synthesizer. With this azobenzene-tethered DNA, formation and dissociation of DNA duplex can be reversibly photo-regulated by cis-trans isomerization of the azobenzene. When the azobenzene takes trans-form, a stable duplex is formed. Upon isomerization of the trans-azobenzene to its cis-form by UV-light irradiation (300 nm < l < 400 nm), the duplex can be dissociated to two strands. The duplex is re-formed on photo-induced cis→trans isomerization (l > 400 nm). By introducing azobenzenes into T7 promoter at specific positions, transcription by T7-RNA polymerase is also efficiently and reversibly photo-regulated. The reversible regulation can be repeated many times without causing damage to DNA or the azobenzene moiety. All these procedures take about 10 days to complete.

  214. Construction of novel dye aggregates based on "comb-type" sequence

    Hiromu Kashida, Taiga Fujii, Hiroyuki Asanuma

    Nucleic Acids Symp. Ser.   51 巻   頁: 11-12   2007年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語  

    Novel dye aggregates (“comb-type" aggregates) was prepared by hybridizing modified oligodeoxyribo-nucleotides (ODNs), in which dyes were introduced consecutively. From NMR structural analysis, dye molecules were intercalated between base pairs and stacked in anti-parallel manner. When ODNs containing three Methyl Red moieties were hybridized, strong exciton coupling was observed. In addition, thermal stability of duplex was substantially enhanced due to the intermolecular stacking.

  215. Enhancement of RNA cleavage activity of 10-23 DNAzyme by covalently introduced intercalator 査読有り

    H. Asanuma, H. Hayashi, J. Zhao, X. Liang, A. Yamazawa, T. Kuramochi, D. Matsunaga, Y. Aiba, H. Kashida, M. Komiyama

    Chem. Commun.     頁: 5062-5064   2006年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    By introducing an intercalator through D-threoninol to the 10-23 DNAzyme at the junction between its catalytic loop and the binding arm, the RNA cleavage activity was greatly improved.

  216. Azobenzene-tethered T7 promoter for Efficient Photoregulation of Transcription 査読有り

    Minghe Liu, Hiroyuki Asanuma, Makoto Komiyama

    J. Am. Chem. Soc.   128 巻   頁: 1009-1015   2006年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Azobenzene was additionally introduced into side chain of T7 promoter for the photoregulation of transcription reaction by T7 RNA polymerase (T7 RNAP). When single azobenzene molecule was introduced into the T7 promoter either at the loop binding region of the RNAP (–7 to –11 position) or at the unwinding region (+1 to –4 position), transcription was suppressed in the trans-form but proceeded faster in the cis-form. The amount of transcripts after UV irradiation with respect to that under dark was only 1.5 – 2.0-fold. Kinetic analysis of the transcription reaction revealed that photoregulatory mechanism was different in these positions. The photoisomerization of an azobenzene at the loop binding region primarily affected Km. On the other hand, the isomerization of an azobenzene at unwinding region mainly affected kcat. Still more clear-cut photoregulation was achieved when two azobenzenes were introduced into both loop binding and unwinding regions, respectively: transcription proceeded 7.6-fold faster after UV irradiation than that under dark. This synergistic effect was observed only when two azobenzenes were introduced into these two different regions, respectively, and introduction into the same loop binding region drastically lowered the transcription activity. The cooperation of two azobenzenes at loop binding and unwinding region would contribute to the clear-cut photoregulation of transcription.

  217. Importance of the Position of Vinyl Group on b-Cyclodextrin for the Effective Imprinting of Amino Acid Derivatives and Oligopeptides in Water 査読有り

    Osawa, T.; Shirasaka, K.; Matsui, T.; Akiyama, T.; Yoshihara, S.; Hishiya, T.; Asanuma, H.; Komiyama, M.

    Macromolecules   39 巻   頁: 2460-2466   2006年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Two kinds of vinyl monomers of b-cyclodextrin (b-CyD) that tether a vinyl group on either wider rim of the truncated cone or its smaller rim were synthesized and applied to the imprinting towards amino acid derivatives and oligopeptides in water. Mono-3-(N-acrylamido)-3-deoxy-altro-b-cyclodextrin (3-AAm-CyD) showed remarkable imprinting effect for the enantioselective recognition of protected amino acids such as N-benzyloxycarbonyltyrosine (Z-Tyr). However, the imprinted polymer from mono-6-(N-acrylamido)-6-deoxy-b-cyclodextrin (6-AAm-CyD) hardly showed enantioselectivity. According to NOESY analysis on pre-organized b-CyD/Z-Tyr complex in D2O, the aromatic moieties of Z-Tyr were included into the cavity of b-CyD from its wider rim. Since the vinyl group of 3-AAm-CyD protruded towards the template and was polymerized there, detailed shape of the template was precisely copied on the polymer by the imprinting. In case of 6-AAm-CyD, however, the shape of template could not be well transcribed because its vinyl group was located at opposite side of the cavity and thus co-polymerization occurred far from the template molecule. On the other hand, the imprinted polymers from both b-CyD vinyl monomers were effective for the recognition of sequences of tetrapeptides composed of two glycines and two phenylalanines, although the selectivity itself was not remarkable. In these polymers, even the b-CyD residues of 6-AAm-CyD were immobilized complementarily to the phenyl rings and bound them.

  218. Improved Method of Molecular Imprinting of Cyclodextrin on Silica-gel Surface for the Preparation of Stable Stationary HPLC Phase 査読有り

    Matsui, T. , Osawa , T.; Shirasaka , K.; Katayama, M.; Hishiya, T.; Asanuma H., Komiyama, M.

    J. Inclu. Phenom. Macrocyclic Chem.   56 巻   頁: 39-44   2006年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    By using N-(3-triethoxysilyl)propylacrylamide (TPAAm), vinyl groups were introduced onto the surface of silicagel.
    On the surface of this silica-gel, b-CyD was molecularly imprinted by using a redox initiator, and the composite
    was used as stationary phase of high performance liquid chromatography (HPLC). The pump pressure was sufficiently
    low and did not increase even after continuous elution for 24 h. In order to prepare still more stable columns,
    a new polymerization process was developed. There, the redox initiator was first mixed with the surface-modified
    silica-gel and then vinylated b-CyD, crosslinker, and the template were added. This modification promoted the
    immobilization of b-CyD copolymer to the silica-gel, resulting in still lower pump pressure. Concurrently, the
    imprinting efficiency was increased in comparison with previous method where the redox initiator was directly
    added to the mixture of the b-CyD–template complex, crosslinker, and surface-modified silica-gel. The molecularly
    imprinted b-CyD column, prepared by this new method, efficiently discriminated the enantiomers of
    N-benzyloxycarbonyltyrosine.

  219. Exciplex Formation between Pyrene and N,N-Dimethylaniline in DNA for the Detection of One-base Deletion 査読有り

    Hiromu Kashida, Makoto Komiyama, Hiroyuki Asanuma

    Chem. Lett.   35 巻 ( 8 ) 頁: 934-935   2006年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Exciplex emission from pyrene and N,N-dimethylaniline moieties was observed in aqueous solution by incorporating them into DNA. By using exciplex formation, one-base deletion was successfully detected.

  220. Activation of DNA enzyme 10-23 by tethering an intercalator to its backbone

    H. Hayashi, X. Liang, J. Zhao, M. Komiyama, and H. Asanuma

    Nucleic Acids Res. Supple   50 巻   頁: 167-168   2006年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語  

  221. Insertion of Two Pyrene Moieties to Oligodeoxyribonucleotides for the Efficient Detection of Insertion/Deletion Polymorphisms 査読有り

    Hiromu Kashida, Hiroyuki Asanuma, Makoto Komiyama

    Chem. Commun.     頁: 2768-2770   2006年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    For the detection of deletion polymorphisms, two pyrene moieties are tethered to oligodeoxyribonucleotide (ODN) on both sides of the intervening base. When this probe ODN was hybridized with wild type ODN, both pyrenes intercalated between base pairs and only monomer emission was observed. In contrast, strong excimer emission was generated by hybridization with one-base deletion mutant. Similarly, two-base deletion was also detected.

  222. Covalent Incorporation of Methyl Red Dyes into Double-Stranded DNA for Their Ordered Clustering 査読有り

    Kashida, H.; Tanaka, M.; Baba, S.; Sakamoto, T.; Kawai, G.; Asanuma, H.; Komiyama, M.

    Chem. Eur. J.   12 巻   頁: 777-784   2006年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Ordered dye cluster of Methyl Reds was formed in double-stranded DNA by hybridizing two complementary DNA-dye conjugates that involve Methyl Red moiety on threoninol linker and 1,3-propanediol as a spacer alternately in the middle of the sequence. In the duplex, Methyl Reds from each strand were axially stacked anti-parallel to each other as determined from NMR analysis. This clustering of Methyl Reds induced distinct change of both UV-Vis and CD spectra. Single-stranded DNA-Methyl Red conjugate on D-threoninol linkers and (1,3-propanediol) spacers exhibited broad absorption spectrum having lmax at around 480 nm, and almost no CD was observed at around absorption maximum of Methyl Red. However, when Methyl Reds were clustered by hybridization, lmax shifted towards shorter wavelength with respect to its monomeric transition. This hypsochromic shift increased with the number of Methyl Reds. Furthermore, positive couplet was also strongly induced here. These dye clusters are H-aggregates, in which molecular excitons are coupled. The positive couplet demonstrates that the clusters on D-threoninol forms right-hand helix. In contrast, induced CD became much weaker with Methyl Red on L-threoninol, which intrinsically prefers counterclockwise winding. Thus, mutual orientation of the stacked dyes was controlled by the chirality of the linker.

  223. Incorporation of methyl group on azobenzene for the effective photo-regulation of hybridization and suppression of thermal isomerization

    H. Nishioka, H. Kashida, M. Komiyama, X. Liang, and H. Asanuma

    Nucleic Acids Res. Supple.   50 巻   頁: 85-86   2006年

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    担当区分:筆頭著者   記述言語:英語  

  224. Clear-cut photo-regulation of the formation and dissociation of the DNA duplex by modified oligonucleotide involving multiple azobenzenes

    Hiroyuki Asanuma, Daijiro Matsunaga, Makoto Komiyama

    Nucleic Acids Res. Supple.   49 巻   頁: 35-36   2005年

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    担当区分:筆頭著者   記述言語:英語  

  225. Design of Light-switchable Phage Promoter for Efficient Photo-regulation of Gene-expression

    Minghe Liu, Hiroyuki Asanuma, Makoto Komiyama

    Nucleic Acids Res. Supple.   49 巻   頁: 283-284   2005年

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    担当区分:筆頭著者   記述言語:英語  

  226. Real time monitoring of the interaction of T7 RNA polymerase with azobenzene-tethered T7 promoter by biosensor

    Minghe Liu, Hiroyuki Asanuma, Makoto Komiyama

    Nucleic Acids Res. Supple. 4   4 巻   頁: 221-222   2004年

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    担当区分:筆頭著者   記述言語:英語  

  227. Photoregulation of in vitro transcription/translation of GFP by tethering an azobenzene to T7 promoter

    Jing Zhao, J. Zhou, Minghe Liu, Hiroyuki Asanuma, Makoto Komiyama

    Nucleic Acids Res. Supple. 4   4 巻   頁: 199-120   2004年

     詳細を見る

    担当区分:筆頭著者   記述言語:英語  

  228. Photo-regulation of RNA Digestion by RNase H with Azobenzene-Tethered DNA 査読有り

    Daijiro Matsunaga, Hiroyuki Asanuma, Makoto Komiyama

    J. Am. Chem. Soc.   126 巻 ( 37 ) 頁: 11452-11453   2004年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    RNA digestion by RNase H, which is responsible for the antisense effect, was efficiently photo-regulated by use of the duplex of azobenzene-tethered sense DNA and native antisense DNA. Under dark, RNA digestion was suppressed because antisense DNA was strongly hybridized with azobenzene-tethered sense DNA, and accordingly RNA was isolated. On UV light irradiation, antisense DNA was released from the azobenzene-tethered DNA due to the trans to cis isomerization and hybridized with RNA, which was digested by RNase H.

  229. Efficient Separation of Hydrophobic Biomolecules by Molecularly Imprinted Cyclodextrins 査読有り

    Hishiya, T.; Asanuma, H.; Komiyama, M.

    J. Inclu. Phenom. Macrocyclic Chem.   50 巻   頁: 51-55   2004年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

  230. Interstrand H aggregation of cationic dyes for narrowing the absorption spectra and stabilizing the duplex 査読有り

    Hiromu Kashida, Hiroyuki Asanuma, Makoto Komiyama

    Supramol. Chem.   16 巻 ( 6 ) 頁: 459-464   2004年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Interstrand H-aggregates of cationic dyes are prepared by hybridization of the two DNA-dye conjugates involving Naphthyl Red moiety and spacer alternately in the middle of the sequence. At pH 5.0 where Naphthyl Red is positively charged, single-stranded DNA-Naphthyl Red conjugate involving three dye groups and three spacer residues exhibited broad absorption spectrum having lmax at around 530 nm. But hybridization of two DNA-Naphthyl Red conjugates that are complementary provided rather different spectrum: much narrower absorption band appeared at 507 nm, which is 23 nm shorter than the single stranded conjugates. These spectroscopic behaviors indicate that dyes in the duplex are H-aggregated and excitons are strongly coupled in the aggregate. In addition to the appearance of narrow H-band, melting temperature dramatically increased by the H-aggregation of stacked cationic dyes compared with that of natural duplex without dye and spacer residues. Thus, positive charges on the stacked dyes did not interfere the duplex formation by the electrostatic repulsion, and moreover fairly promoted the hybridization.

  231. Alternating hetero H-aggregation of different dyes by interstrand stacking from two DNA-dye conjugates

    Hiromu Kashida, Hiroyuki Asanuma, Makoto Komiyama

    Angew. Chem. Int. Ed.   43 巻 ( 47 ) 頁: 6522-6525   2004年

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    担当区分:筆頭著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    New hetero aggregates in which Methyl Reds and Naphthyl Reds are stacked alternately were successfully prepared by hybridization of two DNA-dye conjugates. When the Naphthyl Red conjugate was hybridized with Methyl Red conjugate, a new sharp absorption band (H-band) appeared at 480 nm that was different from the lmax of each dye conjugate in the single-stranded state. In addition to this new band in the UV-Vis spectrum, strong circular dichroism was also induced by interstrand hetero-stacking of these dyes. These results demonstrated that even different dyes as well as identical dyes could exhibit H-band.

▼全件表示

書籍等出版物 27

  1. 生体材料化学 : 基礎と応用

    浅沼 浩之, 樫田 啓, 神谷 由紀子

    コロナ社  2015年  ( ISBN:9784339067507

     詳細を見る

    記述言語:日本語

    CiNii Books

  2. 光応答性DNAの搭載によるナノマシンとナノ構造体の光制御

    浅沼浩之, 神谷由紀子( 担当: 共著)

    化学同人  2021年 

     詳細を見る

    記述言語:日本語

  3. DNA機能の拡張 査読有り

    浅沼浩之( 担当: 単著)

    講談社  2020年12月  ( ISBN:978-4-06-520786-4

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    記述言語:日本語 著書種別:学術書

  4. 核酸の直交性

    浅沼浩之、村山恵司( 担当: 共著)

    CBI学会出版  2019年4月  ( ISBN:978-4-9909076-4-8

  5. ナノマテリアルとしての光応答性DNA

    浅沼浩之、神谷由紀子( 担当: 共著)

    エヌ・ティー・エス  2018年12月 

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    総ページ数:806   担当ページ:7   記述言語:日本語 著書種別:学術書

  6. 遺伝子を蛍光検出するナノプローブ開発 査読有り

    浅沼浩之、樫田 啓( 担当: 共著)

    丸善プラネット(株)  2017年12月 

     詳細を見る

    記述言語:日本語

  7. 非環状骨格型人工核酸:aTNA, SNA

    神谷由紀子、村山恵司、樫田 啓、浅沼浩之( 担当: 共著)

    シーエムシー出版  2016年 

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    記述言語:日本語

  8. 非環状骨格型人工核酸:aTNA, SNA

    神谷由紀子, 村山恵司, 樫田 啓, 浅沼浩之( 担当: 共著)

    シーエムシー出版  2016年 

     詳細を見る

    担当ページ:79-86   記述言語:日本語

  9. 生体材料化学-基礎と応用―

    浅沼浩之、樫田啓、神谷由紀子( 担当: 共著)

    (株)コロナ社  2015年11月  ( ISBN:978-4-339-06750-7

     詳細を見る

    記述言語:日本語

  10. プローブへの応用を目指した人工ヌクレオチドの設計

    浅沼浩之、樫田 啓( 担当: 共著)

    一粒書房  2014年 

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    記述言語:日本語

  11. 核酸医薬を目指した機能性siRNA

    神谷由紀子、浅沼浩之( 担当: 共著)

    一粒書房  2014年 

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    記述言語:日本語

  12. ナノ環境に優しい光応答性DNAの設計

    神谷由紀子、浅沼浩之( 担当: 共著)

    コロナ社   2013年 

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    記述言語:日本語

  13. Oligonucleotide Conjugates for Detection of Specific Nucleic Acid Sequences

    Hiromu Kashida, Hiroyuki Asanuma( 担当: 共著)

    RSC publishing   2012年12月 

     詳細を見る

    記述言語:英語

  14. 核酸の配列特異的認識のためのオリゴヌクレオチドコンジュゲート

    樫田 啓, 浅沼 浩之( 担当: 共著)

    the Royal Society of Chemisry  2012年 

     詳細を見る

    記述言語:英語

  15. カートリッジ型人工ヌクレオチドによる光応答性DNAの設計

    浅沼浩之, 梁興国( 担当: 共著)

    化学同人  2011年 

     詳細を見る

    記述言語:日本語

  16. 人工核酸を利用したプローブDNAの調製と遺伝子多型検出

    樫田啓、浅沼浩之( 担当: 共著)

    NTS  2010年4月 

     詳細を見る

    記述言語:日本語

    これまでに様々な蛍光色素で修飾した核酸プローブが合成されており、溶液中での遺伝子多型の検出に関しては数多くの報告がなされてきた。しかしながら、それに比べて基板にこれらの核酸プローブを固定化した例は必ずしも多くない。その原因としては、プローブ自身の蛍光によるバックグラウンドが挙げられる。蛍光色素で修飾した核酸プローブを基板上に固定化する場合、プローブ自身が発する蛍光により検出が阻害されてしまう。そのため、蛍光性核酸プローブを固定化したDNAチップを調製するためには、ターゲット非存在下でプローブ自身の蛍光を抑制することが必要となる。そこで、本節ではこのようなプローブ単体でのバックグラウンドが抑制された核酸プローブの例として1)モレキュラービーコン及び2)レシオメトリックプローブについて述べる。

  17. 人工核酸を利用したプローブDNAの調製と遺伝子多型検出

    樫田啓, 浅沼浩之( 担当: 共著)

    NTS  2010年4月 

     詳細を見る

    担当ページ:603-610   記述言語:日本語

    これまでに様々な蛍光色素で修飾した核酸プローブが合成されており、溶液中での遺伝子多型の検出に関しては数多くの報告がなされてきた。しかしながら、それに比べて基板にこれらの核酸プローブを固定化した例は必ずしも多くない。その原因としては、プローブ自身の蛍光によるバックグラウンドが挙げられる。蛍光色素で修飾した核酸プローブを基板上に固定化する場合、プローブ自身が発する蛍光により検出が阻害されてしまう。そのため、蛍光性核酸プローブを固定化したDNAチップを調製するためには、ターゲット非存在下でプローブ自身の蛍光を抑制することが必要となる。そこで、本節ではこのようなプローブ単体でのバックグラウンドが抑制された核酸プローブの例として1)モレキュラービーコン及び2)レシオメトリックプローブについて述べる。

  18. DNAのダイナミックな光制御(超分子サイエンスー基礎から材料への展開ー

    浅沼浩之、梁興国( 担当: 共著)

    NTS  2009年5月 

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    記述言語:日本語

    天然のDNAの更なる機能化を目的としてこれまでに様々な化学修飾がDNAに対して行われてきたが、上述のようにDNAの応用分野が多様化したことで、DNAの化学修飾の重要性は更に増しつつある。もちろん天然の4つの核酸塩基だけでも多彩なナノ構造を形成し様々な機能を発揮するが、化学修飾によって天然のDNAでは実現不可能な機能を補うことが出来れば、DNAの応用範囲は飛躍的に拡大するであろう。本稿ではこのような観点から筆者らが開発したアゾベンゼン導入型光応答性DNAと、これを用いたDNAナノマシンのダイナミックな光制御について解説したい。

  19. 光機能化DNAファイバー(”ファイバー”スーパーバイオミメティックス)

    浅沼浩之( 担当: 単著)

    NTS  2006年10月 

     詳細を見る

    記述言語:日本語

  20. 光による核酸機能の制御(図解 高分子新素材の全て)

    浅沼浩之( 担当: 単著)

    工業調査会  2005年 

     詳細を見る

    記述言語:日本語

  21. 光による核酸機能の制御(図解 高分子新素材の全て)

    浅沼浩之( 担当: 単著)

    工業調査会  2005年 

     詳細を見る

    担当ページ:50-53   記述言語:日本語

  22. Molecular imprinting – from fundamentals to applications-

    Makoto Komiyama, Toshifumi Takeuchi, Takashi Mukawa, Hiroyuki Asanuma( 担当: 共著)

    Wiley-VCH  2003年 

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    記述言語:英語

  23. Molecular imprinting – from fundamentals to applications-

    Makoto Komiyama, Toshifumi Takeuchi, Takashi Mukawa, Hiroyuki Asanuma( 担当: 共著)

    Wiley-VCH  2003年 

     詳細を見る

    担当ページ:47-64, 119-138   記述言語:英語

  24. Encyclopedia of Separation Science

    Naoki Toshima, and Hiroyuki Asanuma( 担当: 共著)

    Academic Press Ltd.  2000年 

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    記述言語:英語

  25. Synthesis of Hybrid Polymers Which Bind Guests in Water by Hydrogen Bond Formation

    Makoto Komiyama and Hiroyuki Asanuma( 担当: 共著)

    2000年 

     詳細を見る

    記述言語:英語

  26. Polymer for Gas Separation

    Naoki Toshima, Hiroyuki Asanuma( 担当: 共著)

    VHC Publishers  1992年 

     詳細を見る

    記述言語:英語

  27. Polymer for Gas Separation

    Naoki Toshima, Hiroyuki Asanuma( 担当: 共著)

    VHC Publishers  1992年 

     詳細を見る

    担当ページ:147   記述言語:英語

▼全件表示

MISC 21

  1. DNA-assisted swarm control in a biomolecular motor system

    Keya J., Suzuki R., Kabir A., Inoue D., Asanuma H., Sada K., Hess H., Kuzuya A., Kakugo A.  

    Nature Communications9 巻 ( 1 ) 頁: 453   2018年12月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:Nature Communications  

    In nature, swarming behavior has evolved repeatedly among motile organisms because it confers a variety of beneficial emergent properties. These include improved information gathering, protection from predators, and resource utilization. Some organisms, e.g., locusts, switch between solitary and swarm behavior in response to external stimuli. Aspects of swarming behavior have been demonstrated for motile supramolecular systems composed of biomolecular motors and cytoskeletal filaments, where cross-linkers induce large scale organization. The capabilities of such supramolecular systems may be further extended if the swarming behavior can be programmed and controlled. Here, we demonstrate that the swarming of DNA-functionalized microtubules (MTs) propelled by surface-adhered kinesin motors can be programmed and reversibly regulated by DNA signals. Emergent swarm behavior, such as translational and circular motion, can be selected by tuning the MT stiffness. Photoresponsive DNA containing azobenzene groups enables switching between solitary and swarm behavior in response to stimulation with visible or ultraviolet light.

    DOI: 10.1038/s41467-017-02778-5

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  2. Development of Visible-Light-Responsive RNA Scissors Based on a 10–23 DNAzyme

    Kamiya Y., Arimura Y., Ooi H., Kato K., Liang X., Asanuma H.  

    ChemBioChem19 巻 ( 12 ) 頁: 1305 - 1311   2018年6月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ChemBioChem  

    The 10–23 DNAzyme is an artificially developed functional oligonucleotide that can cleave RNA in a sequence-specific manner. In this study, we designed a new photo-driven DNAzyme incorporating a photoresponsive DNA overhang complementary to the catalytic core region. The photoresponsive overhang region of the DNAzyme included either azobenzene components (Azos) or 2,6-dimethyl-4-(methylthio)azobenzene units (SDM-Azos) each attached to a d-threoninol linker. When the Azos or SDM-Azos were in the trans form, the photoresponsive DNA overhang hybridized with the DNAzyme, and the RNA cleavage activity was suppressed. cis Isomerization of Azos or SDM-Azos, induced by 365 or 400 nm light, respectively, destabilized the duplex between the photoresponsive overhang and the catalytic core, and the DNAzyme recovered RNA cleavage activity. Reversible photoswitching of the DNAzyme activity was achieved by use of specific light irradiation. Further, light-dependent photoswitching of protein expression in the presence of the DNAzyme was demonstrated. Thus, this photo-driven DNAzyme has potential for application as a photocontrolled gene silencing system and a photoactivatable gene expression system.

    DOI: 10.1002/cbic.201800020

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  3. DNA Microcapsule for Photo-Triggered Drug Release Systems

    Kamiya Y., Yamada Y., Muro T., Matsuura K., Asanuma H.  

    ChemMedChem12 巻 ( 24 ) 頁: 2016 - 2021   2017年12月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ChemMedChem  

    In this study we constructed spherical photo-responsive microcapsules composed of three photo-switchable DNA strands. These strands first formed a three-way junction (TWJ) motif that further self-assembled to form microspheres through hybridization of the sticky-end regions of each branch. To serve as the photo-switch, multiple unmodified azobenzene (Azo) or 2,6-dimethyl-4-(methylthio)azobenzene (SDM-Azo) were introduced into the sticky-end regions via a d-threoninol linker. The DNA capsule structure deformed upon trans-to-cis isomerization of Azo or SDM-Azo induced by specific light irradiation. In addition, photo-triggered release of encapsulated small molecules from the DNA microcapsule was successfully achieved. Moreover, we demonstrated that photo-triggered release of doxorubicin caused cytotoxicity to cultured cells. This biocompatible photo-responsive microcapsule has potential application as a photo-controlled drug-release system.

    DOI: 10.1002/cmdc.201700512

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  4. Introduction of 2,6-Diaminopurines into Serinol Nucleic Acid Improves Anti-miRNA Performance

    Kamiya Y., Donoshita Y., Kamimoto H., Murayama K., Ariyoshi J., Asanuma H.  

    ChemBioChem18 巻 ( 19 ) 頁: 1917 - 1922   2017年10月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ChemBioChem  

    MicroRNAs (miRNAs) are endogenous small RNAs that regulate gene expression at the post-transcriptional level by sequence-specific hybridisation. Anti-miRNA oligonucleotides (AMOs) are inhibitors of miRNA activity. Chemical modification of AMOs is required to increase binding affinity and stability in serum and cells. In this study, we synthesised AMOs with our original acyclic nucleic acid, serinol nucleic acid (SNA), backbone and with the artificial nucleobase 2,6-diaminopurine. The AMO composed of only SNA had strong nuclease resistance and blocked endogenous miRNA activity. A significant improvement in anti-miRNA activity of the AMO was achieved by introduction of a 2,6-diaminopurine residues into the SNA backbone. In addition, we found that the enhancement in AMO activity depended on the position of the 2,6-diaminopurine residue in the sequence. The high potency of the SNA-AMOs suggests that these oligomers will be useful as therapeutic reagents for control of miRNA function in patients and as tools for investigating the roles of microRNAs in cells.

    DOI: 10.1002/cbic.201700272

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  5. Chaperone-Polymer-Assisted, Photodriven DNA Strand Displacement

    Cheng Bohao, Kashida Hiromu, Shimada Naohiko, Maruyama Atsushi, Asanuma Hiroyuki  

    CHEMBIOCHEM18 巻 ( 16 ) 頁: 1568-1572 - 1572   2017年8月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ChemBioChem  

    Photodriven DNA strand displacement by using a 2′,6′-dimethylazobenzene-tethered strand and poly(l-lysine)-graft-dextran (PLL-g-Dex) as a chaperone is reported. Rapid strand displacement was reversibly induced by UV and visible-light irradiation without any toehold portion. To further improve the method, the concentration of PLL-g-Dex and the number of equivalents of the photoresponsive strand were optimised. Optimally, 64 % strand displacement was reversibly induced by alternating UV and visible-light irradiation.

    DOI: 10.1002/cbic.201700202

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  6. D-aTNA Circuit Orthogonal to DNA Can Be Operated by RNA Input via SNA

    Murayama K., Nagao R., Asanuma H.  

    ChemistrySelect2 巻 ( 20 ) 頁: 5624 - 5627   2017年7月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ChemistrySelect  

    For a signal amplification system that is orthogonal to DNA, we designed a simplified seesaw gate composed of only D-aTNA. This new system performed signal amplification by toehold exchange reaction just as the DNA circuit did. Moreover, the D-aTNA circuit was not affected by natural nucleic acids carrying sequences complementary to the D-aTNA. In the presence of an SNA interface, however, an RNA signal was converted to D-aTNA signal, resulting in successful activation of D-aTNA circuit. This system can be used to design signal-amplification circuits that are not influenced by contaminating DNA and RNA.

    DOI: 10.1002/slct.201701126

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  7. Orientation-dependent FRET system reveals differences in structures and flexibilities of nicked and gapped DNA duplexes

    Kashida H., Kurihara A., Kawai H., Asanuma H.  

    Nucleic Acids Research45 巻 ( 11 ) 頁: e105   2017年6月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:Nucleic Acids Research  

    Differences in structures and flexibilities of DNA duplexes play important roles on recognition by DNAbinding proteins. We herein describe a novel method for structural analyses of DNA duplexes by using orientation dependence of Forster resonance energy transfer (FRET). We first analyzed canonical B-form duplex and correct structural parameters were obtained. The experimental FRET efficiencies were in excellent agreement with values theoretically calculated by using determined parameters. We then investigated DNA duplexes with nick and gaps, which are key intermediates in DNA repair systems. Effects of gap size on structures and flexibilities were successfully revealed. Since our method is facile and sensitive, it could be widely used to analyze DNA structures containing damages and non-natural molecules.

    DOI: 10.1093/nar/gkx200

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  8. Design of photofunctional oligonucleotides by copolymerization of natural nucleobases with base surrogates prepared from acyclic scaffolds 招待有り 査読有り

    Asanuma H., Murayama K., Kamiya Y., Kashida H.  

    Polymer Journal49 巻 ( 3 ) 頁: 279 - 289   2017年3月

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    記述言語:英語   掲載種別:記事・総説・解説・論説等(学術雑誌)   出版者・発行元:Polymer Journal  

    Further development of DNA nanotechnology requires new functional oligonucleotides composed of nucleobases beyond the native four. In this review, we demonstrate new methodology for DNA and RNA functionalization using a base surrogate prepared from d-threoninol (2-amino-1,3-butanediol). Using this nucleobase surrogate, we can introduce functional molecules at any position of the sequence. Our methodology is conceptually similar to the copolymerization of multiple monomers: phosphoramidite monomers corresponding to the base surrogate and natural nucleotides are copolymerized on a solid support to prepare the functional oligonucleotides. Copolymerization allows for stable functional motifs, including wedges, interstrand-wedges, dimers and clusters. By selecting suitable functional molecules and motifs, we can design photofunctional oligonucleotides, such as: (1) photoresponsive DNA that enables reversible formation and dissociation of the duplex by photoirradiation; (2) [2+2] photocycloaddition of stilbene derivatives; (3) orientation-dependent FRET (fluorescence (Förster) resonance energy transfer) systems; (4) sequence-specific fluorescent probe for the detection of DNA and RNA; and (5) functional siRNA for fluorescent labeling of mature RISC (RNA-induced silencing complex).

    DOI: 10.1038/pj.2016.120

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  9. Effect of Methyl Group on Acyclic Serinol Scaffold for Tethering Dyes on the DNA Duplex Stability

    Murayama K., Asanuma H.  

    ChemBioChem18 巻 ( 1 ) 頁: 142 - 149   2017年1月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ChemBioChem  

    Acyclic serinol derivatives are useful scaffolds for tethering dyes within DNA duplexes. Here we synthesised an inverse l-threoninol (il-threoninol) scaffold and compared its effect on DNA duplex stability to other acyclic artificial nucleic acid scaffolds that are based on d-threoninol, l-threoninol, and serinol. When planar trans-azobenzene was incorporated into the DNA duplex through a single bulge-like motif (the wedge), the il-threoninol scaffold stabilised the duplex most efficiently. When scaffolds were incorporated in complementary positions (dimer motif) or in three adjacent positions (cluster motif), d-threoninol was the most stabilising. CD spectra indicated that the effect of scaffold on the duplex stability was closely related to the winding induced by each scaffold. When trans-azobenzene was photo-isomerised to non-planar cis-azobenzene, il-threoninol destabilised the duplex most strongly, irrespective of the number of artificial residues incorporated. The properties of the il-threoninol scaffold make it a useful tether for dyes or other functionalities.

    DOI: 10.1002/cbic.201600558

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  10. Antisense oligonucleotide modified with serinol nucleic acid (SNA) induces exon skipping in mdx myotubes 査読有り

    Bao T. Le, Keiji Murayama, Fazel Shabanpoor, Hiroyuki Asanuma, Rakesh N. Veedu  

    RSC ADVANCES7 巻 ( 54 ) 頁: 34049 - 34052   2017年

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ROYAL SOC CHEMISTRY  

    Serinol nucleic acid (SNA) is a novel nucleic acid analogue that can form highly stable heteroduplexes with complementary DNA and RNA sequences. Structurally, SNA is a close mimic to peptide nucleic acid (PNA) which is widely used in diagnostic and therapeutic applications. SNA chemistry is relatively new, and so far the scope of SNA has only been explored in improving the efficacy of small interfering RNA and for developing a highly sensitive molecular beacon for diagnostic applications. In this study, we investigated the potential of SNA-modified antisense oligonucleotide (AO) in parallel to PNA-oligo for splicemodulation in an in vitro cellular model of Duchenne muscular dystrophy (DMD). We synthesized a 20mer SNA and PNA antisense oligonucleotide (AO) designed to induce exon-23 skipping in the mouse dystrophin gene transcript. Our results demonstrated that the SNA AO induced exon-23 skipping at all tested concentrations, whereas the corresponding PNA AO failed to induce any exon-23 skipping upon 24 hours of transfection using Lipofectin transfection reagent. Our results further expands the potential of SNA oligonucleotides in therapeutic applications.

    DOI: 10.1039/c7ra06091b

    Web of Science

  11. Antisense oligonucleotide modified with serinol nucleic acid (SNA) induces exon skipping in mdx myotubes

    Bao T. Le, Keiji Murayama, Fazel Shabanpoor, Hiroyuki Asanuma, Rakesh N. Veedu  

    RSC ADVANCES7 巻 ( 54 ) 頁: 34049 - 34052   2017年

     詳細を見る

    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ROYAL SOC CHEMISTRY  

    Serinol nucleic acid (SNA) is a novel nucleic acid analogue that can form highly stable heteroduplexes with complementary DNA and RNA sequences. Structurally, SNA is a close mimic to peptide nucleic acid (PNA) which is widely used in diagnostic and therapeutic applications. SNA chemistry is relatively new, and so far the scope of SNA has only been explored in improving the efficacy of small interfering RNA and for developing a highly sensitive molecular beacon for diagnostic applications. In this study, we investigated the potential of SNA-modified antisense oligonucleotide (AO) in parallel to PNA-oligo for splicemodulation in an in vitro cellular model of Duchenne muscular dystrophy (DMD). We synthesized a 20mer SNA and PNA antisense oligonucleotide (AO) designed to induce exon-23 skipping in the mouse dystrophin gene transcript. Our results demonstrated that the SNA AO induced exon-23 skipping at all tested concentrations, whereas the corresponding PNA AO failed to induce any exon-23 skipping upon 24 hours of transfection using Lipofectin transfection reagent. Our results further expands the potential of SNA oligonucleotides in therapeutic applications.

    DOI: 10.1039/c7ra06091b

    Web of Science

  12. Hetero-Selective DNA-Like Duplex Stabilized by Donor-Acceptor Interactions 査読有り

    Doi T., Sakakibara T., Kashida H., Araki Y., Wada T., Asanuma H.  

    Chemistry - A European Journal21 巻 ( 45 ) 頁: 15974 - +   2015年11月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:Chemistry - A European Journal  

    We report on the characterization of a novel hetero-selective DNA-like duplex of pyrene and anthraquinone pseudo base pairs. The pyrene/anthraquinone pairs showed excellent selectivity in hetero-recognition and even trimers were found to form a hetero-duplex. Pyrene and anthraquinone moieties were tethered on acyclic D-threoninol linkers and linked to adjacent residues by using standard phosphoramidite chemistry. When pyrene and anthraquinone were incorporated at pairing positions in complementary strands of natural DNA oligonucleotides, the duplex was stabilized significantly. Moreover, a pyrene hexamer and an anthraquinone hexamer formed a stable artificial hetero-duplex without the assistance of natural base pairs. The pyrene/anthraquinone pair was so stable that even trimers formed a hetero-duplex under conditions in which natural DNA strands of three residues do not.

    DOI: 10.1002/chem.201502653

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  13. Ultrasensitive molecular beacon designed with totally serinol nucleic acid (SNA) for monitoring mRNA in cells 招待有り 査読有り

    Murayama K., Kamiya Y., Kashida H., Asanuma H.  

    ChemBioChem16 巻 ( 9 ) 頁: 1298 - 1301   2015年6月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:ChemBioChem  

    An artificial nucleic acid based on acyclic serinol building blocks and termed "serinol nucleic acid" (SNA) was used to construct a fluorescent probe for RNA visualization in cells. The molecular beacon (MB) composed of only SNA with a fluorophore at one terminus and a quencher at the other was resistant to enzymatic digestion, due to its unnatural acyclic scaffold. The SNA-MB could detect its complementary RNA with extremely high sensitivity; the signal-to-background (S/B) ratio was as high as 930 when perylene and anthraquinone were used as the fluorophore and quencher pair. A high S/B ratio was also achieved with SNA-MB tethering the conventional Cy3 fluorophore, and this probe enabled selective visualization of target mRNA in fixed cells. Thus, SNA-MB has potential for use as a biological tool capable of visualizing RNA in living cells. Real-time imaging of RNA in living cells: Molecular beacons synthesized from an artificial nucleic acid, serinol nucleic acid (SNA), have extremely high sensitivity for target mRNA. Exogenous and endogenous transcripts were selectively visualized in cells. Thus, SNA is a promising artificial nucleic acid for manipulating RNA in living cells.

    DOI: 10.1002/cbic.201500167

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  14. Evaluation of intrinsic spectroscopic properties of chromophore assemblies by shielding with cyclohexyl base pairs within a DNA duplex 査読有り

    Kashida H., Higashiyama N., Kato T., Asanuma H.  

    Bioorganic and Medicinal Chemistry21 巻 ( 20 ) 頁: 6191 - 6197   2013年10月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:Bioorganic and Medicinal Chemistry  

    Here, we investigated spectroscopic behaviors of tetramethylrhodamine (TMR) homo- and hetero-dimers within DNA duplex. In order to shield the chromophores from natural base pairs, we used cyclohexyl base pairs as 'insulators'; these pairs were inserted between the chromophores and nucleobases. When a single TMR moiety was sandwiched between cyclohexyl base pairs, the emission intensity increased by fivefold relative to a TMR between natural base pairs, because electron transfer from nucleobases was suppressed. Next, we inserted two TMRs between the cyclohexyl base pairs and found that they facilitated H-dimer formation of TMR; a distinct hypsochromic shift was induced only when cyclohexyl base pairs were inserted. We further examined quenching behavior of a TMR paired with a quencher dye between cyclohexyl base pairs. Interestingly, fluorescence from TMR was quenched by nitro methyl red more efficiently in the presence of cyclohexyl base pairs than in their absence. This suggests that neighboring natural base pairs disturbed electron or hole transfer between the fluorophore and the quencher. The cyclohexyl base pairs shielded the chromophore pair from the natural base pairs and allowed intrinsic electron transfer.

    DOI: 10.1016/j.bmc.2013.04.032

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  15. Highly stable duplex formation by artificial nucleic acids aTNA and SNA with acyclic scaffolds 査読有り

    Murayama, K, Tanaka, Y, Toda, T, Kashida, H, Asanuma, H  

    Chem. Eur. J.19 巻 ( 42 ) 頁: 14151 - 14158   2013年

     詳細を見る

    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:Chemistry - A European Journal  

    The stabilities of duplexes formed by strands of novel artificial nucleic acids composed of acyclic threoninol nucleic acid (aTNA) and serinol nucleic acid (SNA) building blocks were compared with duplexes formed by the acyclic glycol nucleic acid (GNA), peptide nucleic acid (PNA), and native DNA and RNA. All acyclic nucleic acid homoduplexes examined in this study had significantly higher thermal stability than DNA and RNA duplexes. Melting temperatures of homoduplexes were in the order of aTNA > PNA ≈ GNA ≥ SNA << RNA > DNA. Thermodynamic analyses revealed that high stabilities of duplexes formed by aTNA and SNA were due to large enthalpy changes upon formation of duplexes compared with DNA and RNA duplexes. The higher stability of the aTNA homoduplex than the SNA duplex was attributed to the less flexible backbone due to the methyl group of D-threoninol on aTNA, which induced clockwise winding. Unlike aTNA, the more flexible SNA was able to cross-hybridize with RNA and DNA. Similarly, the SNA/PNA hetero-duplex was more stable than the aTNA/PNA duplex. A 15-mer SNA/RNA was more stable than an RNA/DNA duplex of the same sequence. © 2013 Wiley-VCH Verlag GmbH & Co. KGaA.

    DOI: 10.1002/chem.201301578

    Scopus

    PubMed

  16. Selective labeling of mature RISC using a siRNA carrying fluorophore–quencher pair 査読有り

    Kamiya Y., Ito A., Ito H., Urushihara M., Takai J., Fujii T., Liang X., Kashida H., Asanuma H.  

    Chemical Science4 巻 ( 10 ) 頁: 4016 - 4021   2013年

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:Chemical Science  

    RNA interference (RNAi) is an endogenous gene silencing system that has been harnessed to inhibit expression of specific genes through the introduction of short double-stranded RNAs, called small interfering RNAs (siRNAs) into cells. After entry into a cell, an siRNA is assembled into the RNA-induced silencing complex (RISC) and suppresses the target gene translation through interaction between the antisense (guide) strand of the siRNA and the target mRNA. To evaluate the intracellular fate of siRNAs, we performed imaging analyses using siRNAs labeled with newly designed fluorophore–quencher clusters introduced through the base surrogate with d-threoninol as a scaffold. Fluorescence microscopy analyses indicated that mature RISC containing fluorescently labeled antisense strand was localized within P-bodies. Thus, the antisense strand uptake into the RISC machinery was successfully visualized. © 2013 The Royal Society of Chemistry.

    DOI: 10.1039/c3sc51197a

    Web of Science

    Scopus

  17. Ultrafast photoisomerization and its single-shot pump pulse efficiency of trans-azobenzene derivative: Compound for photosensitive DNA 査読有り

    Chen T., Yamaguchi A., Igarashi K., Nakagawa N., Nishioka H., Asanuma H., Yamashita M.  

    Optics Communications285 巻 ( 6 ) 頁: 1206 - 1211   2012年3月

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    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)   出版者・発行元:Optics Communications  

    The femtosecond photoisomerization processes of trans (T) 4-carboxy-2′,6′-dimethylazobenzen, which has been employed recently as an efficient photoregulator of DNA hybridization, were clarified by the rate equation analysis of measured transient absorbance changes with (350 nm) and without (380 nm) ground-state absorption of both the reactant (T) and photoproduct (cis: C) isomers under S 2T-band excitation (360 nm, 150 fs pump): after excitation to the S 2T state with a 450-fs lifetime, ∼ 1.5% of the T-molecules in the S 2T state are isomerized to the C-form within ∼ 6 ps through the intermediate state (so called bottleneck state), but most of those return back to the T ground-state S 2T via the internal conversion processes with an ultrafast kinetic rate of 2.2 × 10 12 s - 1. Moreover, the rate equation analysis enables us to determine the T-to-C photoisomerization rate η T,C per pump pulse to be 0.0011 at the pump energy of 80 nJ from the amplitude A 3,350 of the offset component in the 350-nm probe signal, and to obtain the photoisomerization quantum yield Φ T,C = 0.094. The latter value is slightly lower than that of T-azobenzene, and well agrees with that (Φ T,C = 0.097) measured by the conventional CW irradiation method using a photostationary state. © 2011 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.optcom.2011.10.032

    Web of Science

    Scopus

  18. Nick Sealing by T4 DNA Ligase on a Modified DNA Template Tethering a Functional Molecule on D-Threoninol 査読有り

    梁興国, 藤岡健太, 浅沼浩之  

    Chem. Eur. J.17 巻   頁: 10388-10396   2011年

     詳細を見る

    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)  

  19. Photon-fueled DNA nanodevice carrying two different photoswitches 査読有り

    西岡英則, 梁興国, 加藤智博, 浅沼浩之  

    Angew. Chem. Int. Ed.   頁: ***-***   2011年

     詳細を見る

    記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)  

  20. DNAを切断するはさみ

    西岡英則, 浅沼浩之  

    化学64 巻   頁: 70-71   2009年

     詳細を見る

    記述言語:日本語   掲載種別:記事・総説・解説・論説等(学術雑誌)  

    遺伝子工学(遺伝子組み換え技術)は、1)遺伝子であるDNAの特定の位置での“裁断”、2)得られたDNA断片の、ベクターと呼ばれる遺伝子の“運び屋”への“縫製”、そして3)ベクターの細胞内への導入(形質転換)、という3つの基本技術に基づいている。この中でも1)のDNAを“裁断”する “はさみ”=制限酵素 の発見が、現在の遺伝子工学を可能にしたと言っても過言ではない。この天然由来の“はさみ”に相当する制限酵素には様々な種類が存在し、その多くは4~6塩基程度の塩基配列を認識してDNAを切断する。したがって数千塩基程度の短い遺伝子ならば、天然の制限酵素を用いることで特定の1~2箇所を切断することが可能である。しかしながら、高等生物のような巨大なゲノムDNAに対して利用すると非常に多くの断片が生じてしまう。例えば、30億塩基を持つヒトゲノムに対して6塩基しか認識しない制限酵素を利用すると、おおよそ73万 (= 30億÷46)もの断片が生じてし

  21. Unexpected efficient ab initio DNA synthesis at low temperature by using thermophilic DNA

    梁興国, 加藤智博, 浅沼浩之  

    Nucleic Acids Symp. Ser.51 巻   頁: 351-352   2007年11月

     詳細を見る

    記述言語:英語   掲載種別:研究発表ペーパー・要旨(全国大会,その他学術会議)  

    DNA was synthesized in the absence of DNA or RNA as template (or primer) from dNTPs at relatively low temperatures (25~50oC) by thermophilic Vent DNA polymerase whose proper reaction temperature for primer extension is 70~80oC. Unexpectedly, the ab initio DNA synthesis was even more efficient at 50oC as compared with that at 70oC. Interestingly, the ab initio DNA synthesis by Vent (exo-), a mutant version of Vent DNA polymerase lacking of 3&amp;cent;&amp;reg;5&amp;cent; exonuclease activity, became much less efficient, and it could only carry out ab intio DNA synthesis after a long incubation time. This remarkable difference between Vent and Vent (exo-) indicates that the exonuclease activity domain of Vent may play an important role at the initiation step of ab initio DNA synthesis.

▼全件表示

講演・口頭発表等 98

  1. RNAより前に、XNAの世界は存在したか? 招待有り

    浅沼浩之

    生命の起原および進化学会 シンポジウム w/ABC早稲田サテライト  2021年3月11日  生命の起原および進化学会

     詳細を見る

    開催年月日: 2021年3月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    開催地:On-line   国名:日本国  

  2. 非環状型人工核酸SNAとその類縁体の医療展開 招待有り

    浅沼浩之

    令和2年度東海シンポジウム  2021年1月15日  高分子学会東海支部

     詳細を見る

    開催年月日: 2021年1月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    開催地:On-line   国名:日本国  

  3. How far can we escape from DNA? 招待有り 国際会議

    Hiroyuki Asanuma

    Program Conference CLiC digital summer school  

     詳細を見る

    開催年月日: 2020年8月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Fraukfurt on line   国名:日本国  

  4. Photoresponsive XNAs for the photo-switching of bio- and nano-functions 国際会議

    Hiroyuki Asanuma

    China-Japan Joint Interdisciplinary Symposium on Molecular Magnetic Materials 

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    開催年月日: 2019年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Guangzhou   国名:中華人民共和国  

  5. Acyclic XNAs of controlled orthogonality towards DNA and RNA 招待有り

    Hiroyuki Asanuma

    The Fourth A3 Roundtable Meeting on Asia Chemical Probe Research Hub  

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    開催年月日: 2019年11月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  6. Acyclic SNA and aTNA as a new class of XNA for bio- and nanotechnology 招待有り

    Hiroyuki Asanuma

    ISNAC 2019  

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    開催年月日: 2019年10月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Koganei Civic Center   国名:日本国  

  7. DNA nanocapsule for photo-triggered drug release 招待有り 国際会議

    Hiroyuki Asanuma

    2019 International Research Forum on Biology and Medical Science (IRFBMS 2019)  

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    開催年月日: 2019年8月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:台湾  

  8. DNA and XNA nanomachines powered by light irradiation 招待有り

    Hiroyuki Asanuma

    10th NTTH Joint Symposium  

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    開催年月日: 2019年7月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Hotel Hokke Club Hakodate   国名:日本国  

  9. Design of functional oligonucleotide with acyclic scaffold 招待有り 国際会議

    Hiroyuki Asanuma

     詳細を見る

    開催年月日: 2019年6月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Shenyang Pharmaceutical University   国名:中華人民共和国  

  10. 非環状型人工核酸が拓く新たなバイオテクノロジー 招待有り

    浅沼 浩之

    ワークショップ「核酸化学の開く未来」 

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    開催年月日: 2019年6月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  11. Azobenzene-tethered DNA for photo-triggered nanocapsule for drug release 招待有り 国際会議

    Hiroyuki Asanuma

    World Chemistry Conference and Exhibition 2019 (WCCE-2019) 

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    開催年月日: 2019年6月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Brussels, Belgium   国名:ベルギー王国  

  12. Totally acyclic XNAs for medical applications 招待有り 国際会議

    Hiroyuki Asanuma

    Functional Nucleic Acids: From Laboratory to Targeted Molecular Therapy(FNA Perth 2018) 

     詳細を見る

    開催年月日: 2018年11月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:オーストラリア連邦  

  13. DNA二重鎖のナノフォトニクスへの応用 招待有り

    浅沼 浩之

    第22回 VBLシンポジウム  

     詳細を見る

    開催年月日: 2018年11月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  14. Azobenzene-Tethered DNA for Photo-Driven Nanomachine 招待有り 国際会議

    Hiroyuki Asanuma

    XIX NOST-Organic Chemistry Conference (XIX-NOST-OCC) 

     詳細を見る

    開催年月日: 2018年9月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Hotel Grand Hyatt, Goa, India   国名:インド  

  15. Light-triggered DNA nanomachine 招待有り

    Hiroyuki Asanuma

    FISNA2018 

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    開催年月日: 2018年7月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  16. 医療応用を目指した核酸の機能的再インストール 招待有り

    浅沼浩之

    第29回万有仙台シンポジウム―未来を指向した有機化学 

     詳細を見る

    開催年月日: 2018年6月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    開催地:仙台   国名:日本国  

  17. 非環状型人工核酸による分子演算の拡張と核酸医薬への展開 招待有り

    浅沼浩之

    情報計算化学生物学会(CBI学会) 

     詳細を見る

    開催年月日: 2018年5月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    開催地:グランフロント大阪 ナレッジキャピタル   国名:日本国  

  18. DNA nanomachine and nanoarchitecture powered by light-irradiation 招待有り 国際会議

    Hiroyuki Asanuma

    The International Conference of Layers, Films and Membranes for Green, Environmental and Biomedical Sciences (LFM2018) 

     詳細を見る

    開催年月日: 2018年5月

    記述言語:英語   会議種別:口頭発表(基調)  

    開催地: National Taiwan University of Science and Technology, Taipei (Tiwan)   国名:台湾  

  19. Creation of functional oligonucleotide with nucleotide-analogues designed from acyclic scaffold 招待有り

    Hiroyuki Asanuma

     詳細を見る

    開催年月日: 2018年3月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  20. Azobenzene-tethered DNA for the photo-regulation of DNA functions 招待有り 国際会議

    Hiroyuki Asanuma

    INTERNATIONAL CONGRESS ON PURE & APPLIED CHEMISTRY 2018 (ICPAC2018) 

     詳細を見る

    開催年月日: 2018年3月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Siem Reap, Cambodia   国名:カンボジア王国  

  21. Light-driven DNA nanomachine with an azobenzene-tethered DNA as photon-engine 国際会議

    Hiroyuki Asanuma

    the Pure and Applied Chemistry International Conference 2018 (PACCON 2018) 

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    開催年月日: 2018年2月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地: Hat Yai, Songkhla (Thailand)   国名:タイ王国  

  22. Light-driven molecular machines with azobenzene-tethered DNA 招待有り 国際会議

    Hiroyuki Asanuma

    International Conference on Natural and Artificial Molecular Machines (NAMM2018) 

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    開催年月日: 2017年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:IIT Bombay (India)   国名:インド  

  23. "Design of totally acyclic XNAs for medical applications 招待有り

    Hiroyuki Asanuma

    The Second International Symposium on Biofunctional Chemistry (ISBC2017) 

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    開催年月日: 2017年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Kyoto Univ. (Kyoto)   国名:日本国  

  24. SNA and iL-aTNA for nucleic acid medicine 招待有り

    Hiroyuki Asanuma

    1st Minisymposium on Material Biology  

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    開催年月日: 2017年10月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Tokyo institute of Technology   国名:日本国  

  25. Orientation-dependent FRET in DNA duplex 招待有り

    Hiroyuki Asanuma

    FISNA2017 

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    開催年月日: 2017年7月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  26. Orientation-Dependent FRET in DNA Duplex for Structural Analysis of Secondary Structure in Solution 招待有り 国際会議

    Hiroyuki Asanuma

    NTTH symposium 

     詳細を見る

    開催年月日: 2017年7月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Takayama, Gifu   国名:日本国  

  27. Programmable Artificial Nucleic Acids for Bio-application 招待有り

    Hiroyuki Asanuma

    The 20th ISIR International Symposium  

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    開催年月日: 2016年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Osaka, Japan   国名:日本国  

  28. D-L-aTNA and SNA as a new class of acyclic XNA 招待有り 国際会議

    Hiroyuki Asanuma

    Beilstein Organic Chemistry Symposium 2016 Nucleic Acid Chemistry  

     詳細を見る

    開催年月日: 2016年10月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地: Prien (Chiemsee), Germany   国名:ドイツ連邦共和国  

  29. Physics in DNA(4); [2+2] photodimerization of stilbene derivatives within the DNA duplex 招待有り 国際会議

    Hiroyuki Asanuma, Tetsuya Doi, Hiromu Kashida

    The First Roundtable Meeting on Chemical Probe Research Hub 

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    開催年月日: 2016年9月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地: Fukuoka, Japan   国名:日本国  

  30. Orientation-dependent FRET in DNA duplex 国際会議

    Hiroyuki Asanuma

    3rd Fluorescent Biomolecules and Their Building Blocks-Design and Applications (FB3) 

     詳細を見る

    開催年月日: 2016年7月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:中華人民共和国  

  31. Tracing the Fate of siRNA

    Hiroyuki Asanuma

    FIBER International Summit for Nucleic Acids 2016 (FISNA2016) 

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    開催年月日: 2016年7月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  32. 非環状型人工核酸による光機能性ナノマテリアルの創製

    浅沼浩之

    第65回高分子学会年次大会 

     詳細を見る

    開催年月日: 2016年5月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    開催地:神戸国際会議場・展示場 神戸   国名:日本国  

  33. Stemless linear probe for the detection of RNA in living cell 国際会議

    H. Asanuma, M. Akahane, R. Niwa, Y. Kamiya, H. Kashida

    Pacifichem2015  

     詳細を見る

    開催年月日: 2015年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:アメリカ合衆国  

  34. Stemless linear probe for detecting mRNA in living cell 国際会議

    Hiroyuki Asanuma, Mariko Akahane, Yukiko Kamiya, Hiromu Kashida

    BIT's 6th World Gene Convention-2015 

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    開催年月日: 2015年11月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:中華人民共和国  

  35. RNA detection in living cell with super-sensitive linear probe 国際会議

    Hiroyuki Asanuma

    International Conference on Small Science 2015 (ICSS-2015) 

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    開催年月日: 2015年11月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:タイ王国  

  36. Tracing the "fate" of siRNA 国際会議

    Yukiko Kamiya, Anna Ito, and Hiroyuki Asanuma

    The 3rd China-Japan Symposium on Nanomedicine 

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    開催年月日: 2015年6月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:中華人民共和国  

  37. Stemless linear probe with multiple fluorophores on D-threoninols for thefluorescent imaging of m-RNA in cell. 国際会議

    Hiroyuki Asanuma, Mariko Akahane, Hiromu Kashida, Yukiko Kamiya

    1st International Caparica Conference on Chromogenic and Emissive Materials 

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    開催年月日: 2014年9月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:ポルトガル共和国  

  38. de novo Design of functional oligonucleotide with acyclic scaffold 国際会議

    Hiroyuki Asanuma

     詳細を見る

    開催年月日: 2014年7月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:グレートブリテン・北アイルランド連合王国(英国)  

  39. Orientation-dependent FRET between the intercalated donor and acceptor fluorophores 国際会議

    Hiromu Kashida, Ayako Kurihara, Tomohiro Kato, and Hiroyuki Asanuma

    XVIth Symposium on Chemistry of Nucleic Acid Components 

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    開催年月日: 2014年6月

    記述言語:英語   会議種別:口頭発表(一般)  

    国名:チェコ共和国  

  40. Design of highly sensitive fluorescent probe prepared from acyclic threoninol 国際会議

    Hiroyuki Asanuma, Yukiko Kamiya Hiromu Kashida, Xingguo Liang

    2014 China-Japan-Korea and Southeast Asia Joint Symposium on ADVANCED PROCESSING TECHNOLOGY and SAFETY CONTROL of AQUATIC PRODUCTS 

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    開催年月日: 2014年5月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:中華人民共和国  

  41. Functional siRNA for RNAi activation, improvement of strand selection, and fluorescence tracing of RISC 国際会議

    Hiroyuki Asanuma

    RNA-Methods Workshop 

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    開催年月日: 2013年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:ドイツ連邦共和国  

  42. Azobenzene as a molecular engine for driving DNA-based nanomachine with light 国際会議

    Hiroyuki Asanuma

    Selective Regulation in Nanoscaled systems 

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    開催年月日: 2013年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:ドイツ連邦共和国  

  43. Orientation-dependent FRET between the fluorophores within DNA duplex 国際会議

    Hiroyuki Asanuma

     詳細を見る

    開催年月日: 2013年11月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:大韓民国  

  44. Functional siRNA for fluorescent monitoring of RISC with improved activity and strand selectivity

    Hiroyuki Asanuma, Anna Ito, Junya Takai, Masaaki Urushihara, Hiroshi Ito, Taiga Fujii, Xingguo Liang, and Yukiko Kamiya

     詳細を見る

    開催年月日: 2013年11月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  45. Physics in DNA 国際会議

    Hiroyuki Asanuma

    A3RONA 2013 in Kobe 

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    開催年月日: 2013年8月 - 2013年9月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  46. DNA nano-engineering for the construction of photon-fuelled molecular-machine with azobenzenes as molecular engine 国際会議

    Hiroyuki Asanuma

    French-Japanese Seminar on Bioinspired Methods and Applications 

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    開催年月日: 2013年2月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  47. DNA nano-engineering for the construction of photon-fuelled molecular-machine.

    H. Asanuma, H. Nishioka, X.G. Liang, Y. Kamiya

    5th NTTH symposium 

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    開催年月日: 2012年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  48. Control of emission property by assembling dyes for designing intelligent probe 国際会議

    Hiroyuki Asanuma

    2012 Telluride Workshop on Nucleic Acid Chemistry 

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    開催年月日: 2012年7月 - 2012年8月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:アメリカ合衆国  

  49. Photon-fuelled DNA nanomachine carrying azobenzene as molecular engine 国際会議

    Hiroyuki Asanuma

    CIMTECH2012 

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    開催年月日: 2012年6月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:イタリア共和国  

  50. Highly Sensitive Fluorescent Probe designed with Threoninol-Nucleotides 国際会議

    Hiroyuki Asanuma

    A3RONA 2012 

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    開催年月日: 2012年5月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:大韓民国  

  51. New Artificial Nucleic Acids from Acyclic Scaffolds

    K. Murayama, T. Toda, H. Kashida, and H. Asanuma

    BMMP-12 

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    開催年月日: 2012年1月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  52. Artificial nucleotides for therapy-siRNA activation and probing-

    Hiroyuki Asanuma

    GCOE satellite sympoium  

     詳細を見る

    開催年月日: 2011年11月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  53. Photon-fuelled nanomachine carrying azobenzene as molecular engine 国際会議

    Hiroyuki Asanuma

    A3RONA2011 

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    開催年月日: 2011年10月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:中華人民共和国  

  54. OLD BUT NEW ARTIFICIAL NUCLEIC ACIDS FROM ACYCLIC THREONINOL (aTNA) AND SERINOL (SNA) 国際会議

    Hiromu Kashida, Keiji Murayama, Takasuke Toda, Xingguo Liang, and Hiroyuki Asanuma

    XVth Symposium on Chemistry of Nucleic Acid Components 

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    開催年月日: 2011年6月

    記述言語:英語   会議種別:口頭発表(基調)  

    国名:チェコ共和国  

  55. Light-up of DNA with artificial nucleotides

    H. Kashida, K. Sekiguchi, H. Asanuma

    BMMP11 

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    開催年月日: 2011年1月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  56. 次世代核酸医薬を目指した人工核酸の設計

    浅沼浩之

    名古屋大学予防早期医療創成センター 第4回研究会 

     詳細を見る

    開催年月日: 2010年11月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  57. Light-up of DNA with Threoninol-Nucleotides 国際会議

    A3RONA 2010 JAPAN 

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    開催年月日: 2010年10月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  58. 人工DNAを活用した簡便かつ高感度検出が可能なプローブ設計

    浅沼浩之

    生体分子計測のボトムアップテクノロジー 

     詳細を見る

    開催年月日: 2010年10月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  59. AZOBENZENE-TETHERED DNA FOR THE PHOTOREGULATION OF DNA FUNCTIONS

    IRT 2010 - XIX International Round Table on Nucleosides, Nucleotides and Nucleic Acids 

     詳細を見る

    開催年月日: 2010年8月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

  60. カートリッジ型人工核酸によるDNAの光機能化

    国際バイオEXPO 

     詳細を見る

    開催年月日: 2010年6月

    記述言語:日本語   会議種別:口頭発表(一般)  

    国名:日本国  

  61. RATIONAL DESIGN OF PHOTORESPONSIVE DNA WITH CARTRIDGE-TYPE NUCLEOTIDE 国際会議

    The 4th International Symposium on Polymer Chemistry (PC2010) 

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    開催年月日: 2010年6月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

  62. Design of in-stem molecular beacon (ISMB) with "threoninol nucleotide" for the discrimination of polymorphisms 国際会議

    Gene-2009 

     詳細を見る

    開催年月日: 2009年12月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  63. 遺伝子発現の光制御を目指した光応答性DNAの開発

    第30回日本レーザー医学会総会 

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    開催年月日: 2009年12月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  64. Threoninol-nucleotidesによるDNAの再インストール

    浅沼浩之

    第58回高分子討論会 

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    開催年月日: 2009年9月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  65. Rational design of functional DNA with threoninol-nucleotides 国際会議

    Vielberth-Symposium on Functional Nucleic Acids 

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    開催年月日: 2009年9月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

  66. Photo-driven DNA Nanomachine with New Duplex Motif Composed of Threoninol 国際会議

    International Conference on Materials for Advanced Technologies 

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    開催年月日: 2009年7月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    In the present paper, we demonstrate photoresponsive DNA tweezers as a nanomachine fuelled with photons by using azobenzene-modified DNA. The tweezers are opened by UV light irradiation (330-350 nm) and closed by visible light irradiation (440-460 nm) without adding oligonucleotides as the fuel.

  67. 光機能性インテリジェントDNAの設計と応用

    平成21年度第三回PST-net 例会 

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    開催年月日: 2009年4月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    光機能性DNAを用いた機能性材料の設計と応用について講演する。

  68. 光が導くバイオテクノロジー

    「光科学技術と応用」シンポジウム 

     詳細を見る

    開催年月日: 2009年2月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    光応答性DNAを用いた遺伝子発現の光制御に関する今後の展望について講演する。

  69. DNAを利用した光機能マテリアルの開発

    平成20年度東海シンポジウム 

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    開催年月日: 2009年1月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    我々は、非環状リンカーのD-Threoninolを活用した機能分子の導入法が上記の目的に合うことを見出した。D-ThreoninolをScaffoldに用いて機能分子(インターカレーター)を導入した“人工塩基=Threoninol nucleotide”は、任意の配列に任意の数DNA中に導入することが可能であり、しかも二重鎖の不安定化や基質特異性を損なわずに二重鎖内にインターカレートする。

  70. Photoregulation of Gene Expression with Azobenzene-tethered Promoter 国際会議

    Ninth International Symposium on biomimetic Materials Processing (BMMP-9) 

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    開催年月日: 2009年1月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    In the present paper, we propose another strategy for the photoregulation of gene expression with the azobenzene-tethered DNA. Azobenzene is introduced into the promoter region (T7-promoter) of RNA polymerase (RNAP) and transcription reaction by RNAP is photoregulated by trans-cis isomerization of the incorporated azobenzenes.

  71. 非環状リンカーによるDNAの再インストール

    日本学術振興会第174委員会第27回研究会 

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    開催年月日: 2008年12月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    本講演でリボースに代わる新たなScaffoldとして非環状リンカーのD-Threoninol(Fig.1a参照)を活用した“DNAの再インストール”と、その応用展開について解説する。

  72. PHOTOREGULATION OF DNA FUNCTIONS BY AZOBENZENE-TETHERED OLIGONUCLEOTIDES 国際会議

    NU-UM Joint Symposium 

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    開催年月日: 2008年11月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    Here, efficient photo-switching of hybridization with this modified DNA and its application to the bio-reaction are introduced.

  73. 配列の差異を検出するインテリジェントDNAプローブの開発

    第39回中部化学関係学協会支部連合秋季大会 

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    開催年月日: 2008年11月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    本講演では、このthreoninol-nucleotideを活用した、SNPsやIndelsを検出可能なインテリジェントDNAプローブについて解説する。

  74. AZOBENZENE-TETHERED PROMOTER TOWARDS THE PHOTO-REGULATION OF GENE EXPRESSION 国際会議

    Japan-Korea Polymer Young Scientists Symposium 

     詳細を見る

    開催年月日: 2008年10月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    A simple and effective approach for introducing azobenzene moieties tethered on D-threoninol to long duplex DNA (gene) was developed. By PCR and enzymatic ligation, the chemically modified part was introduced to the exact position we expected. By using the constructed photoresponsive gene, the GFP expression could be simply switched on and off by light-irradiation. As the photoswitch was equipped in the promoter, this strategy is promising to photoregulate the expression of any gene or functiona

  75. DNAの再インストールによる光デバイス化

    高分子エレクトロニクス研究会 

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    開催年月日: 2008年6月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    我々はリボースに頼らない非天然機能分子の導入法の一つとしてD-threoninolが非常に有効であることを見出し、これをScaffoldに用いたDNAの“再インストール”を行っている。本講演では、アゾベンゼン導入D-threoninolを使用した光デバイスとしてのDNAの再インストールについて紹介する。

  76. Interstrand Clustering of Dyes in the DNA Duplex with Threoninol 国際会議

    Hiroyuki Asanuma, Taiga Fujii, Hiromu Kashida

     詳細を見る

    開催年月日: 2008年1月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  77. リボースに頼らないDNAの光機能化

    浅沼浩之

    機能性分子シンポジウム 

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    開催年月日: 2008年1月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    本発表では、D-threoninolをScaffoldに使用したDNAの再インストールについて、紹介する。

  78. 情報を持った繊維:DNAファイバーの光機能化

    浅沼浩之

    繊維学会夏季セミナー 

     詳細を見る

    開催年月日: 2007年9月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    筆者らは、核酸が関与する酵素反応や分子マシンの光制御を目的とした光機能化DNAファイバーの開発を目指した。光刺激は数ある外部刺激の中でも1)反応系を汚染しない、2)分子設計により励起波長の制御が可能、3)レンズによる集光やレーザー光の利用で局所的な刺激ができる、といった長所を持つ。また近年パルスレーザーの進歩に伴って多光子励起が可能になり、より長波長の光による色素分子の励起も容易になりつつある。ここでは筆者らが近年開発した可逆的な応答を示す光機能化DNAファイバーについて解説する。

  79. DNAの再構築 -リボースを使用しないオリゴヌクレオチドの機能化-

    浅沼浩之

    FIBERシンポジウム 

     詳細を見る

    開催年月日: 2007年6月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    我々は、リボースに頼らない非天然分子の導入法の一つとしてD-threoninolを提案してきた。ScaffoldとしてD-threoninolを用いると、DNA二重鎖の安定性を損なうことなくインターカレーターを多数導入することが出来る。またD-threoninolに導入した機能分子を“擬似塩基”として天然のオリゴヌクレオチド中に導入することも可能である。本講演では、D-threoninolをScaffoldに用いたDNAの機能化について解説する。

  80. 超分子デバイスとしての機能性DNA

    浅沼浩之

    第10回VBLシンポジウム 

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    開催年月日: 2006年10月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    本発表では、筆者らが近年開発した可逆的な応答を示す光デバイス化DNAについて解説する。

  81. Azobenzene-Tethered DNA as a Photo-Switching Biodevice 国際会議

    11th International Symposium on Colloidal and Molecular Electro-Optics (ELOPTO-2006) 

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    開催年月日: 2006年5月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    DNA has not only a biological role of storing genetic information but also various physical and chemical functions. These role and functions are mainly based on the spontaneous hybridization of two strands that are complementary each other. If the formation and dissociation of the DNA duplex can be photo-regulated as schematically illustrated in Fig.1, we can get a new effective photo-switching biodevice and scope of the application should be extended, such as artificial regulation of gene exp

  82. 色素とのコンジュゲーションによるDNAの光機能化

    浅沼浩之

    表面技術協会 第113講演大会 

     詳細を見る

    開催年月日: 2006年3月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    色素分子は、発光・吸収などの光学特性や、特定波長の光照射による可逆的な構造異性化など特異的な性質を有しており、DNAとのコンジュゲーションによって光応答性などDNAに新たな機能の付与が期待できる。逆にDNAの超分子性を活用することで色素分子のクラスター化が容易となり、単量体では実現不可能な光学物性の発現が期待できる。本講演では、互いの特性を生かした色素とDNAの新たなコンジュゲートについて、筆者らの最近の成果を紹介する。

  83. PHOTOREGULATION OF DNA FUNCTIONS BY AZOBENZENE-TETHERED OLIGONUCLEOTIDES 国際会議

    Sixth International Symposium on Biomimetic Materials Processing (BMMP-6) 

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    開催年月日: 2006年1月

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    We have synthesized modified DNA tethering azobenzenes, and hybridization with its complementary strand has been successfully photo-regulated. Here, efficient photo-switching of hybridization with this modified DNA and its application to the bio-reaction are introduced.

  84. 生体機能の光制御を目指したアゾベンゼン導入DNAの設計

    浅沼浩之

    第16回MRSシンポジウム 

     詳細を見る

    開催年月日: 2005年12月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

  85. 機能分子との交互コンジュゲーションが広げるDNAの可能性

    浅沼浩之

    SORST ジョイントシンポジウム (4)  ― クロスオーバーする生命と化学 ― 

     詳細を見る

    開催年月日: 2005年11月

    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    国名:日本国  

    機能分子の核酸への導入は、天然に無い性能をDNAやRNAに付与する上で有効な手段である。しかし化学修飾は多くの場合二重鎖の不安定化や配列特異性の低下を伴うため、5’末端への機能分子の導入が一般的である。もちろん精緻にデザインした化学修飾ヌクレオシドを合成してDNA鎖の内部に導入した例も多数報告されているが、合成や分子設計に煩雑さを伴う場合もある。従ってDNA二重鎖の安定性や配列特異性を損なわず簡便に機能分子を多数導入できる一般的な手法が開発されれば、応用の可能性が大きく広がると期待できる。我々はアゾベンゼン

  86. New Artificial Nucleic Acids from Acyclic Scaffolds 国際会議

    K. Murayama, T. Toda, H. Kashida, H. Asanuma

    BMMP-12  2012年1月24日 

     詳細を見る

    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:名古屋大学  

  87. 非環状型人工核酸が拓く新たなバイオテクノロジー 招待有り 国際会議

    浅沼 浩之

    ワークショップ「核酸化学の開く未来」  2019年6月19日  埼玉大学先端産業国際ラボラトリー

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    記述言語:日本語   会議種別:口頭発表(招待・特別)  

  88. 医療応用を目指した核酸の機能的再インストール 招待有り 国際会議

    浅沼浩之

    第29回万有仙台シンポジウム―未来を指向した有機化学  2018年6月8日  万有財団

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    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    開催地:仙台  

  89. Tracing the "fate" of siRNA

    Yukiko Kamiya, Anna Ito, Hiroyuki Asanuma

    The 3rd China-Japan Symposium on Nanomedicine  2015年6月19日 

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    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Beijing, China  

  90. Totally acyclic XNAs for medical applications 招待有り

    Hiroyuki Asanuma

    Functional Nucleic Acids: From Laboratory to Targeted Molecular Therapy(FNA Perth 2018)  2018年11月22日 

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    記述言語:英語   会議種別:口頭発表(招待・特別)  

  91. Physics in DNA(4); [2+2] photodimerization of stilbene derivatives within the DNA duplex 招待有り

    Hiroyuki Asanuma, Tetsuya Doi, Hiromu Kashida

    The First Roundtable Meeting on Chemical Probe Research Hub  2016年9月22日 

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    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Fukuoka, Japan  

  92. Physics in DNA

    Hiroyuki Asanuma

    A3RONA 2013 in Kobe  2013年8月30日 

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    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:神戸  

  93. Photoresponsive XNAs for the photo-switching of bio- and nano-functions

    Hiroyuki Asanuma

    China-Japan Joint Interdisciplinary Symposium on Molecular Magnetic Materials  2019年12月27日 

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    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Guangzhou  

  94. Photon-fuelled nanomachine carrying azobenzene as molecular engine

    Hiroyuki Asanuma

    A3RONA2011  2011年10月14日 

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    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Wuhan, China  

  95. Orientation-Dependent FRET in DNA Duplex for Structural Analysis of Secondary Structure in Solution 招待有り

    Hiroyuki Asanuma

    NTTH symposium  2017年7月14日 

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    記述言語:英語   会議種別:口頭発表(招待・特別)  

    開催地:Takayama, Gifu  

  96. Orientation-dependent FRET between the intercalated donor and acceptor fluorophores

    Hiromu Kashida, Ayako Kurihara, Tomohiro Kato, Hiroyuki Asanuma

    XVIth Symposium on Chemistry of Nucleic Acid Components  2014年6月8日 

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    記述言語:英語   会議種別:口頭発表(一般)  

    開催地:Český Krumlov  

  97. Orientation-dependent FRET between the fluorophores within DNA duplex

    Hiroyuki Asanuma

    The 9th Korea-Japan Symposium on Frontier Photoscience-2013 (KJFP-2013)  2013年11月24日 

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    記述言語:英語   会議種別:口頭発表(招待・特別)  

  98. OLD BUT NEW ARTIFICIAL NUCLEIC ACIDS FROM ACYCLIC THREONINOL (aTNA) AND SERINOL (SNA)

    Hiromu Kashida, Keiji Murayama, Takasuke Toda, Xingguo Liang, Hiroyuki Asanuma

    XVth Symposium on Chemistry of Nucleic Acid Components  2011年6月5日 

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    記述言語:英語   会議種別:口頭発表(基調)  

▼全件表示

共同研究・競争的資金等の研究課題 3

  1. 非環状型機能性人工核酸の開発

    研究課題番号:AS2915131U  2017年10月 - 2020年3月

    研究成果展開事業 研究成果最適展開支援プログラムA-STEP 

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    配分額:51350000円 ( 直接経費:39500000円 、 間接経費:11850000円 )

  2. DNAハイブリダイゼーションの高効率可逆光スイッチング技術の開発と,そのバイオテクノロジーへの応用

    2005年10月 - 2011年3月

      詳細を見る

    資金種別:競争的資金

  3. 生体反応の光制御を目指した人工核酸デバイスの創製

    2002年11月 - 2006年3月

    科学技術振興機構PRESTO 

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    資金種別:競争的資金

科研費 26

  1. 非環状型人工核酸による人工遺伝システムの創成とその進化分子工学への応用

    研究課題/研究課題番号:21H05025  2021年7月 - 2026年3月

    科学研究費助成事業  基盤研究(S)

    浅沼 浩之, 根本 直人

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:190450000円 ( 直接経費:146500000円 、 間接経費:43950000円 )

    本申請では、1)L-aTNAを”ゲノム”に見立てた自己複製(L-aTNA→L-aTNA)・転写(L-aTNA→DNA(RNA))・逆転写(DNA(RNA)→L-aTNA)という人工遺伝システムを非酵素的に実現することで、L-aTNAがPre-RNAワールド仮説の候補になりうる原始核酸であることを実証し、さらに2)この人工遺伝システムを活用したL-aTNA人工アプタマー創成へと展開する。

  2. 核酸医薬への応用を目指した非環状型人工核酸の開発

    2019年9月 - 現在

    国立研究開発法人日本医療研究開発機構  先端的バイオ創薬等基盤技術開発事業 

    浅沼浩之

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    配分額:146900000円 ( 直接経費:113000000円 、 間接経費:33900000円 )

  3. 直交性人工核酸を用いた、誤作動の無いロバストなシグナル増幅回路の設計

    研究課題/研究課題番号:18H03933  2018年4月 - 2021年3月

    科学研究費助成事業  基盤研究(A)

    浅沼 浩之

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    配分額:44980000円 ( 直接経費:34600000円 、 間接経費:10380000円 )

    D-aTNAは、天然のDNAやRNAとは二重鎖を形成しない。それに対し、我々が開発した人工核酸SNAは、これら3つの核酸と配列特異的に二重鎖を組むことができる。そこでこれらの関係を利用し、SNAをインターフェースに使用して、天然のRNA入力で、これと直交するD-aTNAのみで構成されるシグナル増幅回路が起動するシステムを構築した。こうして入力RNAとクロストークしない直交核酸でシグナル増幅回路を設計・構築し、ヌクレアーゼおよび夾雑DNAやRNAの存在する環境下でも微量のマイクロRNA(miR21)をロバストに検出可能なシグナル増幅回路を実現した。
    本研究が実現したことで、細胞内で微量のRNAをロバストに検出可能な新たな蛍光プローブの設計が可能になり、miRNAを標的とした病理診断など診断薬の開発が期待できる。またこれまの核酸の直交性の概念は天然のDNAあるいはRNAの配列特異性のみに留まっていたが、人工核酸が加わったことで天然および人工核酸間での二重鎖形成能に基づく直交性へと、直交性の概念が大きく拡張された。

  4. 非環状型機能性人工核酸の開発

    研究課題/研究課題番号:AS2915131U  2017年10月 - 2020年3月

    国立研究開発法人科学技術振興機構  研究成果最適展開支援プログラム(A-STEP) 

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    配分額:51350000円 ( 直接経費:39500000円 、 間接経費:11850000円 )

  5. 直交性核酸を使用したクロストーク型増幅回路の設計

    研究課題/研究課題番号:16K12522  2016年4月 - 2018年3月

    科学研究費助成事業  挑戦的萌芽研究

    浅沼 浩之

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    配分額:3510000円 ( 直接経費:2700000円 、 間接経費:810000円 )

    直交性とは、互いに干渉(相互作用)しない一連の要素群の関係を示す。我々は、これまでにセリノール誘導体を用いた3種類の非環状骨格型人工核酸D-aTNA, L-aTNA, SNAの開発に成功している。これらは全て、極めて安定なホモ二重鎖を形成する。D-aTNAは天然のDNAやRNAと二重鎖を形成しないのに対し、アキラルな骨格を持つSNAは、D-, L-aTNA、DNA、RNA全てと安定な二重鎖を形成する。そこで本研究では、RNAと直交しているD-aTNAのみで論理回路を設計し、SNAをインターフェースに用いて、RNAが入力するとD-aTNA 回路が作動するシステムを設計した。

  6. ステム構造を必要としない高感度リニアプローブの創成

    研究課題/研究課題番号:25248037  2013年4月 - 2016年3月

    科学研究費助成事業  基盤研究(A)

    浅沼 浩之, 神谷 由紀子, 樫田 啓

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    配分額:47450000円 ( 直接経費:36500000円 、 間接経費:10950000円 )

    本研究では、ステム構造を必要としないリニアプローブのコンセプトを更に発展させ、消光色素アントラキノンと蛍光色素ペリレンを併用することで超高感度(S/B比1600)と高い酵素耐性を実現し、細胞内でのmRNAの蛍光イメージングを実現した。次にストランドインベージョンを実現するため、蛍光色素としてエチニルペリレンを使用しアントラキノンと共に多数導入したリニアプローブを設計した。設計通りDNAと安定な二重鎖を形成し、PNAとは二重鎖を形成しなかった。その結果、PCR産物でもストランドインベージョンによる二重鎖DNAの直接蛍光ラベルを実現した。

  7. ステム構造を必要としない高感度リニアプローブの創成

    2013年4月 - 2015年3月

    科学研究費補助金  基盤研究(A)

    浅沼浩之

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    配分額:47450000円 ( 直接経費:36500000円 、 間接経費:10950000円 )

    モレキュラービーコン(MB)は汎用性の高い蛍光プローブとして知られているが、ステム・ループ構造に起因する様々な欠点を持つ。我々は、疑似ヌクレオチド化した蛍光色素をDNA 中に適切な間隔で導入することで、ステム・ループ構造を必要としない全く新たなコンセプトの蛍光プローブ“リニアプローブ”の開発に成功した。本申請研究では、リニアプローブのコンセプトを更に発展させて、モレキュラービーコンを遥かに凌駕する高輝度・高感度リニアプローブを開発し、
    1) 高い酵素耐性と高速応答を活かしたm-RNA の細胞内でのイメージング、2) PNA を組み合わせることでストランドインベージョンによる二重鎖DNA の直接蛍光ラベル への応用 を目指す。

  8. 構造化ゲルと化学反応場の協働による運動創発

    研究課題/研究課題番号:24104005  2012年6月 - 2017年3月

    科学研究費助成事業  新学術領域研究(研究領域提案型)

    萩谷 昌己, 村田 智, 浅沼 浩之, 菅原 研, 有村 隆志, 宮元 展義, 原 雄介, 濱田 省吾, 川又 生吹, 浜田 省吾

      詳細を見る

    担当区分:研究分担者  資金種別:競争的資金

    スライム型ロボットの研究では,分子ロボットの「スケールの拡大」を目指した.具体的には,高分子ゲルを反応場として,ミリメートル・オーダーの非均質な反応空間を生成し,その中でさまざまな分子デバイス群を動作させる.このコンセプトにより,最終的には,走性のような機能を目指している.得られた成果としては,分子デバイスのゲル内での反応/拡散の挙動の定量的な測定に基づいてそれを制御する方法,各種の新しいゲルアクチュエータの開発,ゾル・ゲル相転移を制御する方法,ゲルのセル空間上に定義された離散的な反応場をプログラムするためのゲルオートマトンと呼ばれる理論的枠組みなどがあげられる.

  9. 人工核酸を用いた新規ストランドインベーダーの開発

    研究課題/研究課題番号:23651128  2011年 - 2012年

    科学研究費助成事業  挑戦的萌芽研究

    浅沼 浩之

      詳細を見る

    担当区分:研究代表者  資金種別:競争的資金

    配分額:4030000円 ( 直接経費:3100000円 、 間接経費:930000円 )

    本研究では、D-Threoninol に色素などの機能性分子を導入したヌクレオチド・アナログ=Threoninol-Nucleotide (TN)を導入した DNA(TN-DNA)と、通常の未修飾のPNA を組み合わせた新たなストランドインベーダーの開発を目指した。DNA 二重鎖へのインベージョン活性を示すためには、二重鎖の安定性の序列が DNA/PNA, TN-DNA/DNA > DNA/DNA > TN-DNA/PNA を満たす必要がある。そこでこのような安定性の序列を満たすための機能性分子と TN の導入方法を検討した。その結果、電子吸引性基を導入した平面構造のインターカレーターが DNA 二重鎖を大きく安定化することを見出した。また TN を天然のヌクレオチド 2 残基毎に導入すると、PNA の二重鎖を阻害しつつ DNA との二重鎖形成を促進することも明らかにした。これらの結果に基づき、PNA と機能性分子としてアントラキノンを導入した TN-DNA の組み合わせることで、実際に DNA 二重鎖にストランドインベージョンすることを明らかにした。

  10. 光応答性DNAを活用した光駆動型分子マシンの開発

    研究課題/研究課題番号:21241031  2009年 - 2012年

    科学研究費助成事業  基盤研究(A)

    浅沼 浩之, 梁 興国

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:46800000円 ( 直接経費:36000000円 、 間接経費:10800000円 )

    本研究では、アゾベンゼンを導入した光応答性 DNA を用いた光駆動型分子マシンの開発を目指した。まず DNA 二重鎖の形成と解離の高効率な光制御を可能にする修飾アゾベンゼンの分子設計を行った。その結果、主鎖から遠い側にあるベンゼン環の 2 つのオルト位の化学修飾が、光制御効率の向上と cis 体の熱安定性の向上に有効であることを見出した。また二重鎖中でアゾベンゼンと塩基対が交互に並ぶように光応答性 DNA の配列を設計すると、アゾベンゼンの光異性化で二重鎖の形成と解離の完全な光制御が可能なことも見出した。 これらの技術を利用し、 1) RNA を可逆的に切断する光駆動型 DNA 分子マシン、 2) 光崩壊型 DNA ナノカプセル、3) 可視光応答型アゾベンゼンとの組み合わせによるシーソー型ナノマシン の構築に成功した。

  11. スーパー制限酵素による巨大DNAの遺伝子操作

    研究課題/研究課題番号:18001001  2006年 - 2009年

    科学研究費助成事業  特別推進研究

    小宮山 真, 小宮山 真, 浅沼 浩之, 須磨岡 淳

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    担当区分:研究分担者  資金種別:競争的資金

    我々が開発したスーパー制限酵素を使って、ヒトの全ゲノムDNA(30億塩基対)の中の1カ所を選択的に切断することに成功した。DNA切断はすべて天然酵素と同様にリン酸ジエステル結合の加水分解で進行するので、生成したDNA断片を用いて組換えDNAを調製し、これを細胞内で発現することが可能である。さらに、スーパー制限酵素で切断したDNAをヒト細胞内に入れると、この切断がDNA 修復系により正確に認識され、所定の相同組換えが促進されることも明らかにした。

  12. 新規人工核酸"グライコ核酸"の創製

    研究課題/研究課題番号:18655069  2006年 - 2007年

    科学研究費助成事業  萌芽研究

    浅沼 浩之

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:3600000円 ( 直接経費:3600000円 )

    本研究ではペプチド核酸に糖鎖を導入したグライコ核酸を設計・創製する。そして1)糖鎖導入で従来のPNAに両親媒性を付与することで20mer以上の長鎖DNAに対する二重鎖形成能を飛躍的に高める、2)糖鎖の持っ親水性を活用して疎水性の高いインターカレーターの導入を可能にする、ことを目標とする。
    18年度の研究成果において本研究の目標に掲げた親水性の付与を実現したので、19年度は得られたグライコ核酸のタンパク質との特異的相互作用について検討した。まず18年度に確立したグライコ核酸の"ヌクレオシド"モノマーの合成法に従い、アミノ酸リンカーにD-およびL-Lysを用いて機能分子としてマンノースを導入したカートリッジモノマーを設計・合成した。さらにFmoc固相合成法によりマンノースユニットを持った"グライコ核酸"を合成し、マンノースに特異的なレクチンであるConcanavalin A(ConA)を使用して、グライコ核酸との相互作用を蛍光強度の変化で評価した。その結果、PNA中にマンノースユニットを1つ導入したグライコ核酸のConA認識能力は単糖と同程度であったが、二つ導入した系では糖のクラスター効果に由来する高い認識力が認められた。このように、任意の場所に任意の数の糖モノマーを導入できる本研究のグライコ核酸の特徴を活かすことで、高いタンパク質認識能を付与可能なことも明らかとなった。本研究者はアゾベンゼンのような疎水性の高いインターカレーターをPNA中に導入する方法も併せて確立したので、本申請研究のグライコ核酸と組み合わせることで更なる高機能化が期待できる。

  13. 分子ナノマシンの光スイッチングを目指した人工DNAデバイスの構築

    研究課題/研究課題番号:17310066  2005年 - 2007年

    科学研究費助成事業  基盤研究(B)

    浅沼 浩之

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:15530000円 ( 直接経費:14900000円 、 間接経費:630000円 )

    本申請研究では、1)分子ナノマシンの光スイッチングに適した光応答性DNA(=人工DNAデバイス)を開発し、2)光を燃料とする新規な分子ピンセットを構築した。
    分子マシンの"エンジン"部分に相当する光応答性DNAは、精緻な分子設計により、従来型の無置換アゾベンゼンと比較して3倍効率が高い新規なアゾベンゼンの合成に成功した。これによってtrans-体でより二重鎖を安定化し、cis-体で大きく不安定化する新規な光応答性DNAが得られた。また更に精密な分子設計によって、これまでとは逆のスイッチング-trans-体で二重鎖が解離しcis-体で二重鎖を形成する-にも成功した。
    上記で開発した一部の光応答性DNAを活用することで、光駆動型の分子マシン-分子ピンセット-を開発した。従来の天然のDNAを使用した分子ピンセットでは、ピンセット開閉の駆動力としてDNA自身を使用するのに対し、本申請研究の光駆動型分子ピンセットは、光応答性DNAを"エンジン"として搭載している。いわばソーラー型分子ピンセットであり、設計どおり可視光照射でピンセットを閉じUV光照射で開くことが出来た。
    DNA自身を燃料としている従来型の分子ピンセットは、開閉操作の繰り返しでDNA二重鎖が老廃物として不可避的に系内に蓄積した。そのためにピンセットの開閉を繰り返すと効率が顕著に低下してしまった。一方本申請研究で開発した光駆動型分子ピンセットは、"環境に優しく"系内を汚染しない"光"を駆動力に使用しているので、開閉操作を繰り返しても効率の低下は観察されなかった。すなわち本申請研究の分子マシンは、DNAが光分解しない限り半永久的に開閉操作を繰り返すことが可能な分子マシンと言える。

  14. 形態変化する分子を用いた並行計算と分散計算

    研究課題/研究課題番号:14085202  2002年 - 2006年

    科学研究費助成事業  特定領域研究

    萩谷 昌己, 横山 茂之, 陶山 明, 浅沼 浩之, 藤井 輝夫, JOHN Rose, 村田 智, 岩崎 裕, 吉信 達夫

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    担当区分:研究分担者  資金種別:競争的資金

    形態変化するDNA分子の設計方法:萩谷は、連続ヘアピンからなる分子マシンに関して、ヘアピンの配列を変化させたときに、3連続ヘアピンから成る分子マシンの挙動がどのように変化するかを調べた。分子マシンの光制御を目指した光機能性超分子の構築:浅沼は、これまでとは逆に「cis-体で二重鎖形成、trans-体で解離」というスイッチングが可能な光応答性DNAの設計と実現に成功した。ヘアピンとバルジによる並行計算:萩谷は、Whiplash PCRの熱力学的な解析と、状態遷移の効率化(Displacement WPCR)を行った。レトロウィルスによる並行計算:陶山は、二つの正帰還と一つの負帰還反応から構成されたオシレータをRTRACの基本反応を用いて構築した。シミュレーションにより発振可能な条件が存在することを確かめた後、実装を進めた。翻訳系による並行計算:横山は、翻訳システムを利用した「オートマトン」を動物細胞(培養細胞)内でも構築することを目標に、非天然型アミノ酸が細胞に与えられた場合にのみ活性化され、サプレッサーtRNAにアミノ酸を結合するような酵素(アミノアシルtRNA合成酵素)を用いることにより、サプレションを制御する機構を構築した。DNA Walker:萩谷は、DNA Walkerの構築に向けて、温度、pH、光の三種類の入力によって駆動する分子マシンに関する予備実験を行った。特に、これらの三種類の入力の独立性について調べた。マイクロチップのための微量液体制御機構の開発:藤井は、液滴操作に必要な周辺技術等の整備を進め、オンデマンド式で液滴の生成・合一の操作が可能にした。また、本技術を用いてDNAとPNAのハイブリダイゼーション反応と電気泳動による反応産物の分離操作をデバイス上で実現した

  15. 人工制限酵素によるDNAの位置特異的切断と機能性核酸の合成

    研究課題/研究課題番号:13132204  2001年 - 2004年

    科学研究費助成事業  特定領域研究

    小宮山 真, 浅沼 浩之, 須磨岡 淳

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    担当区分:研究分担者  資金種別:競争的資金

    本研究により、DNAおよびRNAの選択的に切断する手法の開発に成功した。
    1.DNA切断
    相補鎖DNAを用いて基質一本鎖DNAにギャップ構造を形成させ、ここにCe(IV)/EDTAを加えると、ギャップ部位での選択的切断が実現することを明らかにした。さらに、相補鎖DNAの末端をリン酸修飾することにより、ギャップでの切断効率が大きく向上することを見出した。
    二本鎖DNAに対してPseudo-complementary PNA (pcPNA)をstrand invasionすることにより基質DNA中に部分的に一本鎖部位を形成させ、ここにCe(IV)/EDTAを加えるとそれぞれの鎖の一本鎖部分での選択的な切断が実現し、結果として二本鎖DNAが塩基配列特異的に切断されることを見出した。さらに、strand invasionさせるpcPNAの末端をリン酸やアミノポリカルボン酸などの配位子で修飾することにより、切断効率が大きく向上することを明らかにした。また、我々の見出した人工制限酵素系によって切断されたDNAは、天然酵素を用いて切断断片と外来DNAを再結合することが可能であり、組換えDNAを調製することにも成功した。
    2.RNA切断
    アクリジンをインターカレーターとしてDNA鎖に導入し、相補的RNAと二本鎖形成させることにより基質RNA中のアクリジンの正面が位置選択的に活性化され、ここに希土類イオンを加えると活性化された部分のみが選択的に切断されるごとを見出した。また、この方法を利用し、RNA中の目的領域を挟むようにDNA鎖に2つのアクリジンを導入し、目的のRNA断片を切り出すことにも成功した。さらに、切断した断片をMALDI-TOF-MSにより質量解析することで、精密かつ高速な一塩基多型の検出法の開発に目途をつけた。

  16. 光応答性DNAによる酵素反応の光制御

    研究課題/研究課題番号:13878116  2001年

    科学研究費助成事業  萌芽的研究

    浅沼 浩之, 小宮山 真

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:2100000円 ( 直接経費:2100000円 )

    本研究では、アゾベンゼンをT7プロモーターに導入し、アゾベンゼンのtrans-cis異性化によるT7-RNAPのプロモーター部位への結合を制御することで、転写反応の光制御を目指した。アゾベンゼンはT7プロモーターのnon-template鎖に導入したところ、プロモーターの上流-10と-11位の間にアゾベンゼンを導入した場合に最も効果的な光制御が実現できた。
    アゾベンゼンの無い天然のプロモーターを用いた場合は、UV照射下でも未照射下とほぼ同じ速度で転写反応が進行した。しかしアゾベンゼン導入プロモーターを用いると、暗条件下では転写産物がほとんど得られなかったのに対し、UV照射下では天然のプロモーターとほぼ同量のm-RNAが得られた。すなわちアゾベンゼンがtrans-体の場合は転写反応が著しく抑制されるのに対し(転写off)、cis-体では転写が天然のプロモーターと同程度進行すると(転写on)が明らかとなった。この光応答性プロモーターを用いれば、転写反応を光照射のみでon-off制御することが可能となった。すなわち、暗条件下では"スイッチ"はoffであったが、UV照射によってスイッチがonになり、m-RNAが生成した。この後、更に可視光照射によって再びスイッチをoffに切り替えることが出来た。この様に、光応答性プロモーターを用いることで転写反応のon-off制御を実現した。

  17. 化学修飾DNAを用いた遺伝子発現の光制御

    研究課題/研究課題番号:11167216  2000年

    科学研究費助成事業  特定領域研究(A)

    浅沼 浩之, 小宮山 真

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:3500000円 ( 直接経費:3500000円 )

    本申請では、遺伝子発現の光制御を目指している。前年度我々は、トリメチレンリンカーの中心の炭素にアゾベンゼンを導入した修飾DNAを合成し、アゾベンゼンの光異性化によって二重鎖の形成と解離を光のみで制御できることを明らかにしている。本年度はその機構を明らかにするため、アゾベンゼンを含むDNAの二重鎖の構造を2D NMRによって解析した。その結果、1)trans-アゾベンゼンが隣接する塩基対間にインターカレートしている、2)trans-アゾベンゼンがインターカレートすることでメジャーグルーブ側に傾いている、ことが判明した。一方cis-体は非平面分子であるため、スタッキングによる安定化が得られずむしろ立体障害で不安定化し、結果としてtrans→cis異性化によって二重鎖形成能が大きく変化することが明らかとなった。また、NOESYの詳細な解析から、極性の高い成分はRのコンフィギュレーションであることも明らかにした。
    更に光応答性DNAを、T7-DNAポリメラーゼによるDNAの伸長反応に応用し、目論見どおりアゾベンゼンの光異性化で、伸長反応をON-OFF制御できることを明らかにした。更に詳細な解析から、モジュレーターが5'末端付近から徐々に剥がれるような機構で光制御が実現できていることを見出した。
    光応答性DNAを酵素反応の制御に応用するためには、天然の構造からの差異は小さいほうが良い。そこでリボースの2'Oにアゾベンゼンを導入したウリジンを合成し、DNA内に組み込んだ。この系は配列依存性が大きいが、trans-cis異性化でTmが変化することを見出した。また、アゾベンゼンと並んで代表的な光異性化分子であるスピロピラン導入DNAも合成に成功した。

  18. モレキュラーインプリント法でシクロデキストリンの配向を制御したレセプターの構築

    研究課題/研究課題番号:11750734  1999年 - 2000年

    科学研究費助成事業  奨励研究(A)

    浅沼 浩之

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:2200000円 ( 直接経費:2200000円 )

    我々はすでに、β-シクロデキストリン(β-CyD)の配向の制御にモレキュラーインプリント法を適用することで、コレステロールを強く認識する架橋シクロデキストリン高分子の合成に成功している。更にMALDI-TOFMSを用いた分析から、コレステロールが鋳型として存在することで、CyDの二量化が促進されることを見出した。また、CyDのビニルモノマーを用いることで水中での鋳型重合を実現し、ジペプチドや抗生物質を選択的に認識する人工レセプターの合成にも成功している。本年度はこの手法の液体高速クロマトグラフィー(HPLC)の担体への応用を目指した。
    バッチテストでインプリント効果が確認されているコレステロールを鋳型として合成した高分子を適当な大きさに粉砕し、ステンレススチールカラムに充填してHPLCとしてのゲスト保持能を検討した。その結果、鋳型分子であるコレステロールに対して高い保持能を示した。一方鋳型分子以外の分子に対する保持能は、わずかしか向上せず、インプリント効果により、鋳型分子に特異的な認識部位が形成されていることが明らかとなった。
    また、メチレンビスアクリルアミドを架橋剤に用いてCyDのビニルモノマーとの共重合により水中で合成した架橋シクロデキストリン高分子は、力学的強度が小さく、耐圧性に乏しかった。そこで、十分な強度を持つシリカゲル上にインプリント高分子を固定化することで耐圧性を向上させることに成功した。このようにして得られたHPLC担体は、水を溶離液として使用しても鋳型分子に対して高い保持能を示すことが明らかとなった。以上の様に、本研究のインプリント高分子は、HPLC担体としても応用可能なことが明らかとなった。

  19. 水系で水素結合により分子認識するハイブリッドポリマーの構築

    研究課題/研究課題番号:10126209  1998年

    科学研究費助成事業  特定領域研究(A)

    浅沼 浩之

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:2000000円 ( 直接経費:2000000円 )

    天然の抗体と同様に水溶液中で基質を正確に認識する人工レセプターの合成は、ホスト・ゲスト化学に携わる研究者にとって究極の目標である。本年度は(1)高分子効果による水溶液中での水素結合形成の促進、ならびに(2)モレキュラーインプリント法によるホスト分子相互の分子配向の制御、を追究した。
    (1) 高分子効果による水溶液中での水素結合形成の促進
    本研究者は、既にポリ(2-ビニル-4,6-ジアミノ-1,3,5-トリアジン)(PVDAT)が水溶液中で、相補的な水素結合によりチミンとウラシルを選択的に認識することを見出している。しかしPVDATは水に不溶の高分子であり、固-液界面という特殊な環境が水中での水素結合に寄与していることも予想された。そこで2-ビニル-4,6-ジアミノ-1,3,5-トリアジン(VDAT)をアクリルアミドと共重合させて水に可溶化し、高分子場が均一水溶液中でも相補的な水素結合形成に有効に機能するか検討した。その結果、上記共重合体は、均一水溶液においても相補的な水素結合を形成してチミンを選択的に認識することが判明した。以上より、高分子場が均一水溶液中でも水素結合形成に有効なことが明らかとなった。
    (2) モレキュラーインプリント法によるホスト分子相互の分子配向の制御
    本研究者は、シクロデキストリン(CyD)を鋳型分子存在下でDMSO中でジイソシアナート架橋することで、水中で基質を選択的に認識する人工レセプターが合成であることを明らかにしている。本年度は、この方法で形成される認識部位の構造を物理化学的な手段を用いて詳細に分析した。その結果、コレステロールを鋳型分子に用いた場合、2分子のβ-CyDがジイソシアナートで架橋された構造を持つ認識部位が高分子内に形成されていることを明らかにした。

  20. 水系で水素結合により分子認識する人工高分子レセプターの開発

    研究課題/研究課題番号:09750964  1998年

    科学研究費助成事業  奨励研究(A)

    浅沼 浩之

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:1800000円 ( 直接経費:1800000円 )

    本研究者は、水に不溶のポリ(2-ビニル-4,6-ジアミノ-1,3,5-トリアジン)(PVDAT)が水溶液からチミンとウラシルを選択的に認識・分離することを見出し、これがジアミノトリアジン残基との相補的水素結合の形成に基づくことを分光学的な手法で明らかにした。高分子場が形成する疎水場は水溶液中だけでなくアルコール中でも有効であり、メタノール中でも相補的水素結合形成によってチミンとウラシルを認識・分離した。またメタノール中ではスタッキング相互作用など疎水相互作用が強く働かないため、相補的水素結合の数に基づく基質選択性は水中よりも高かった。
    さらに高分子場が上記のような不均一系ではなく、均一水溶液系でも有効に疎水場を形成して基質との相補的水素結合が可能となるか検討した。ジアミノトリアジン残基を持つ水溶性高分子は、2-ビニル-4,6-ジアミノ-1,3,5-トリアジンをアクリルアミドと共重合させることで得られた。上記共重合体は、均一水溶中でもチミンを選択的に認識することが、限外ろ過膜を使用した透析実験より判明した。またH-NMR測定より、チミンのイミドプロトンのみが共重合体の存在下で低磁場シフトしたことから、チミンの認識が相補的水素結合に基づいていることを明らかにした。一方対応するモノマーでは、チミンとの相補的水素結合の形成は全く認められなかった。以上より、高分子場が均一水溶液中でも水素結合形成に有効なことが明らかとなった。

  21. 水溶液中で水素結合を形成し、基質を分子認識するハイブリッドポリマーの構築

    研究課題/研究課題番号:09232213  1997年

    科学研究費助成事業  重点領域研究

    小宮山 真, 浅沼 浩之

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    担当区分:研究分担者  資金種別:競争的資金

    これまでに合成されている水素結合部位で分子認識する人工レセプターは、水溶液中では、水分子が水素結合サイトに対して競争阻害を起こすために十分に機能しない。我々は、これらの問題の解決を目指して、高分子効果の活用を検討した。その結果、ジアミノトリアジンを側鎖に持つポリ2-ビニル4,6-ジアミノトリアジン(PVDT)が、水の中でも、相補的な水素結合を効率的に形成し、基質を高選択的に認識することを見出した。すなわち、一連の核酸塩基の中で、ジアミノトリアジンと3本の相補的水素結合を形成するウラシルおよびチミンのみがPVDTにより強く吸着される。それに対して、水素結合サイトが2箇所であるシトシンの吸着はわずかであり、水素結合サイトが1箇所であるピリミジンは吸着されない。このように、PVDTの複合体形成能は、ゲストとの相補的水素結合サイトの数と良好に対応している。PVDTとウラシルとの結合定数をLangmuirプロットにより見積もったところ、93M^<-1>と十分に大きな値であった。それに対して、PVDTの単量体モデルおよび2量体モデルは、ウラシルと水素結合を形成しない。こうして、(1)PVDTが、水溶液中で、水素結合によりゲスト化合物を効率的かつ選択的に分子認識すること、および(2)水中での水素結合形成には高分子場か必須であること が明らかとなった。さらに我々は、目的とするゲスト化合物に対応した人工レセプターを自在に構築する手法の開発を目指して、ゲスト化合物の存在下にシクロデキストリン(CyD)を架橋し、CyD柏互の分子配向を制御した。今年度に主たる認識対象としたのはコレステロールであり、鋳型分子としてのコレステロール存在下でβ‐CyDをDMSO中でジイソシアネート架橋することで、水溶液中でコレステロールを強く認識する人エレセプターの合成にも成功した。

  22. 細胞内微量miRNAの検出を目指した人工核酸によるシグナル増幅回路の開発

    2016年4月 - 2021年3月

    公益財団法人 旭硝子財団  ステップアップ助成 

    浅沼浩之

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:12000000円 ( 直接経費:12000000円 )

  23. 構造化ゲルと化学反応場の協働による運動創発

    2012年10月 - 2017年3月

    科学研究費補助金  研究領域提案型(計画研究)

    浅沼浩之

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    担当区分:研究分担者  資金種別:競争的資金

    配分額:52000000円 ( 直接経費:40000000円 、 間接経費:12000000円 )

  24. 医療応用を目指した人工核酸の創成

    2012年4月 - 2015年3月

    キャノン財団  研究助成プログラム「産業基盤の創生」 

    浅沼浩之

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:20000000円 ( 直接経費:20000000円 )

  25. スーパー制限酵素による巨大DNAの遺伝子操作

    2006年10月 - 2011年3月

    科学研究費補助金 

    小宮山真

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    担当区分:研究分担者 

  26. 新規人工核酸“グライコ核酸”の創製

    2006年

    科学研究費補助金  萌芽研究,課題番号:18655069

    浅沼 浩之

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    担当区分:研究代表者 

▼全件表示

産業財産権 14

  1. 人工核酸を含むオリゴヌクレオチド

    浅沼浩之、樫田啓、村山恵司

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    出願番号:特願2015-003424  出願日:2015年1月

    出願国:国内  

  2. 人工核酸を含むオリゴヌクレオチド

    浅沼浩之, 樫田啓, 村山恵司

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    出願番号:特願2015-003424  出願日:2015年1月

  3. 蛍光標識オリゴヌクレオチド誘導体及びその利用

    浅沼浩之 樫田啓 近藤展代 大澤卓矢 赤羽真理子

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    出願人:名古屋大

    出願番号:特願2011―221332号  出願日:2011年10月

    出願国:国内  

  4. オリゴヌクレオチドプローブ及びその利用

    浅沼浩之、丸山厚、嶋田直彦、梁興国、樫田啓、藤井大雅、大澤卓矢、吉田安子、丹羽孝介

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    出願人:日本碍子株式会社

    出願番号:特願2011-33986  出願日:2011年2月

    出願国:国内  

  5. イン・ステムビーコン型プローブ用のハイブリダイゼーション剤及びその利用

    丸山厚,嶋田直彦,浅沼浩之,梁興国,樫田啓,藤井大雅,大澤卓矢,吉田安子,丹羽孝介

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    出願人:日本碍子株式会社

    出願番号:特願2011―033986号  出願日:2011年

    出願国:国内  

  6. オリゴヌクレオチド及びその利用

    浅沼 浩之、 樫田 啓、 関口 康司、近藤 展代

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    出願番号:特願2010-206043  出願日:2010年9月

    出願国:国内  

  7. オリゴヌクレオチドおよびその利用

    浅沼 浩之、 梁 興国、 樫田 啓、 西岡 英則、藤井 大雅、石川 顕慎

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    出願番号:特願2010-194942  出願日:2010年8月

    出願国:国内  

  8. インスレーター及びその利用

    樫田啓、浅沼浩之、関口康司

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    出願人:法人名大

    出願番号:特願2010-042632  出願日:2010年2月

    出願国:国内  

  9. 標的核酸の検出方法

    浅沼浩之、梁興国、樫田啓、吉田安子、吉良茂樹、丹羽孝介

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    出願人:法人名大、日本ガイシ(株)

    出願番号:特願2009-298170  出願日:2009年12月

    出願国:国内  

  10. オリゴヌクレオチドプローブ及びその利用

    浅沼浩之、梁興国、樫田啓、原雄一、吉田安子、吉良茂樹

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    出願人:法人名大、日本ガイシ(株)

    出願番号:特願2009-298160  出願日:2009年12月

    出願国:国内  

  11. オリゴヌクレオチドプローブ及びその利用

    浅沼浩之、梁興国、樫田啓、吉田安子、高瀬智和、丹羽孝介

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    出願人:法人名大、日本ガイシ(株)

    出願番号:PCT/JP2009/061980  出願日:2009年6月

    出願国:国内  

  12. ターゲットDNAの特異増幅方法の開発

    梁興国、浅沼浩之、鈴木昌友、加藤智博

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    出願番号:特願2009-145741  出願日:2009年6月

    出願国:国内  

  13. オリゴヌクレオチドプローブ及びその利用

    浅沼 浩之、梁 興国、樫田 啓、吉田 安子、高瀬 智和、丹羽 孝介

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    出願番号:特願2008-172526  出願日:2008年7月

    出願国:国内  

  14. DNAエンザイムおよびその活性制御方法

    浅沼浩之、小宮山 真、松永 大次郎、倉持 壮

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    出願人:独立行政法人科学技術振興機構

    出願番号:特願2004-55086  出願日:2005年2月

    公開番号:PCT/JP/2005/003052 

    出願国:外国  

▼全件表示

 

担当経験のある科目 (本学以外) 2

  1. 光スイッチングデバイスとしての光応答性DNAの開発と、その応用

    2007年4月 - 2008年3月 名古屋市立大学 大学院薬学研究科)

  2. 生命分子工学特論

    2005年4月 - 2006年3月 東京大学大学院工学系研究科化学生命工学専攻)