Language：English   Publishing type：Research paper (scientific journal)   Publisher：Genetics Society of America  

Stably transmitted transgenes are indispensable for labeling cellular components and manipulating cellular functions. In Caenorhabditis elegans, transgenes are generally generated as inheritable multi-copy extrachromosomal arrays, which can be stabilized in the genome through a mutagenesis-mediated integration process. Standard methods to integrate extrachromosomal arrays primarily use protocols involving ultraviolet light plus trimethylpsoralen or gamma- or X-ray irradiation, which are laborious and time-consuming. Here, we describe a one-step integration method, following germline-mutagenesis induced by mini Singlet Oxygen Generator (miniSOG). Upon blue light treatment, miniSOG tagged to histone (Histone-miniSOG) generates reactive oxygen species (ROS) and induces heritable mutations, including DNA double-stranded breaks. We demonstrate that we can bypass the need to first establish extrachromosomal transgenic lines by coupling microinjection of desired plasmids with blue light illumination on Histone-miniSOG worms to obtain integrants in the F3 progeny.We consistently obtained more than one integrant from 12 injected animals in two weeks. This optogenetic approach significantly reduces the amount of time and labor for transgene integration. Moreover, it enables to generate stably expressed transgenes that cause toxicity in animal growth.

DOI： 10.1534/g3.118.200158

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Scopus

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELIFE SCIENCES PUBLICATIONS LTD  

Subcellular localization of ribosomes defines the capacity for local translation. Methods to visualize ribosomes in live multicellular organisms are desirable for mechanistic investigations of the cell biology of ribosomes. Here, we developed an approach using split GFP for tissue-specific visualization of ribosomes in live Caonorhabditis elegans. Ribosomes are detected as puncta in the axons and synaptic terminals of specific neuron types, correlating with ribosome distribution at the ultrastructural level. We found that axonal ribosomes change localization during neuronal development and after axonal injury. Using genetic screens, we showed that the microtubule cytoskeleton and the JIP3 protein UNC-16 exert distinct effects on localization of axonal and somatic ribosomes. Our data demonstrate the utility of tissue-specific visualization of ribosomes in vivo, and provide insight into the mechanisms of active regulation of ribosome localization in neurons.

DOI： 10.7554/eLife.26376

Web of Science

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3791/54810

PubMed

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1073/pnas.1514123113

Web of Science

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/ncomms9868

Web of Science

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1534/genetics.114.167841

Web of Science

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.conb.2011.10.002

Web of Science

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1074/jbc.M109.021519

Web of Science

Event date： 2023.9

Language：Japanese   Presentation type：Public lecture, seminar, tutorial, course, or other speech  

Country：Japan  

Event date： 2023.9

Language：Japanese  

Venue：金沢   Country：Japan  

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Venue：名古屋   Country：Japan  

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Venue：名古屋   Country：Japan  

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Event date： 2023.2

Language：English   Presentation type：Poster presentation  

Country：Japan  

Event date： 2022.12

Language：Japanese   Presentation type：Public lecture, seminar, tutorial, course, or other speech  

Country：Japan  

Event date： 2022.11 - 2022.12

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Country：Japan  

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Venue：Online  

Event date： 2022.9

Language：English   Presentation type：Oral presentation (invited, special)  

Venue：Trivandrum   Country：India  

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Venue：Vienna   Country：Austria  

Event date： 2022.6

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Venue：オンライン   Country：Japan  

Event date： 2022.3

Presentation type：Oral presentation (general)  

Event date： 2022.3

Language：Japanese   Presentation type：Public lecture, seminar, tutorial, course, or other speech  

Event date： 2021.7

Language：Japanese   Presentation type：Public lecture, seminar, tutorial, course, or other speech  

Event date： 2021.6

Language：English   Presentation type：Poster presentation