title: Optogenetic Random Mutagenesis Using Histone-miniSOG in C. elegans.
abstract: 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.
authors.affiliation: Division of Biological Sciences, Section of Neurobiology, University of California in San Diego and Howard Hughes Medical Institute; knoma@ucsd.edu.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Noma
authors.affiliation: Division of Biological Sciences, Section of Neurobiology, University of California in San Diego and Howard Hughes Medical Institute; Department of Cellular and Molecular Medicine, University of California in San Diego School of Medicine and Howard Hughes Medical Institute; yijin@ucsd.edu.
authors.forename: Yishi
authors.initials: Y
authors.lastname: Jin
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans; D005854:Germ Cells; D006657:Histones; D016296:Mutagenesis; D062308:Optogenetics; D017382:Reactive Oxygen Species
chemical_list: D006657:Histones; D017382:Reactive Oxygen Species