2016年11月
Optogenetic Random Mutagenesis Using Histone- miniSOG in C. elegans
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
- ,
- 巻
- 号
- 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
- ID情報
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- DOI : 10.3791/54810
- ISSN : 1940-087X
- PubMed ID : 27911392
- Web of Science ID : WOS:000415734600012