論文

2016年11月

Optogenetic Random Mutagenesis Using Histone- miniSOG in C. elegans

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
  • Kentaro Noma
  • ,
  • Yishi Jin

117
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.3791/54810
出版者・発行元
JOURNAL OF VISUALIZED EXPERIMENTS

Forward genetic screening in model organisms is the workhorse to discover functionally important genes and pathways in many biological processes. In most mutagenesis-based screens, researchers have relied on the use of toxic chemicals, carcinogens, or irradiation, which requires designated equipment, safety setup, and/or disposal of hazardous materials. We have developed a simple approach to induce heritable mutations in C. elegans using germline-expressed histone-miniSOG, a light-inducible potent generator of reactive oxygen species. This mutagenesis method is free of toxic chemicals and requires minimal laboratory safety and waste management. The induced DNA modifications include single-nucleotide changes and small deletions, and complement those caused by classical chemical mutagenesis. This methodology can also be used to induce integration of extrachromosomal transgenes. Here, we provide the details of the LED setup and protocols for standard mutagenesis and transgene integration.

リンク情報
DOI
https://doi.org/10.3791/54810
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/27911392
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000415734600012&DestApp=WOS_CPL
ID情報
  • DOI : 10.3791/54810
  • ISSN : 1940-087X
  • PubMed ID : 27911392
  • Web of Science ID : WOS:000415734600012

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