title: Methods for single/low-copy integration by ultraviolet and trimethylpsoralen treatment in Caenorhabditis elegans.
abstract: Single/low-copy transgene integration is essential for avoiding overexpression, ectopic expression and gene silencing in the germline. Here, we present an overview of a method that uses ultraviolet and trimethylpsoralen (UV/TMP) to generate single/low-copy gene integrations in Caenorhabditis elegans. Single/low-copy transgenes from extrachromosomal arrays are integrated into the genome using positive selection based on temperature sensitivity with a vps-45 rescue fragment and negative selection based on benzimidazole sensitivity with a ben-1 rescue fragment. The copy number of the integrated transgenes is determined using quantitative PCR. Our UV/TMP integration method, which is based on familiar extrachromosomal transgenics, provides a simple approach for generating single/low-copy gene integrations.
authors.affiliation: Department of Physiology, Tokyo Women's Medical University, School of Medicine, 8-1, Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan; The OCU Advanced Research Institute for Natural Science and Technology, Osaka City University, 3-3-138, Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan. Electronic address: nakadai@life.osaka-cu.ac.jp.
authors.forename: Eriko
authors.initials: E
authors.lastname: Kage-Nakadai
authors.affiliation: Department of Physiology, Tokyo Women's Medical University, School of Medicine, 8-1, Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. Electronic address: rimae@research.twmu.ac.jp.
authors.forename: Rieko
authors.initials: R
authors.lastname: Imae
authors.affiliation: Department of Physiology, Tokyo Women's Medical University, School of Medicine, 8-1, Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. Electronic address: yoshina@research.twmu.ac.jp.
authors.forename: Sawako
authors.initials: S
authors.lastname: Yoshina
authors.affiliation: Department of Physiology, Tokyo Women's Medical University, School of Medicine, 8-1, Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. Electronic address: mitani1@research.twmu.ac.jp.
authors.forename: Shohei
authors.initials: S
authors.lastname: Mitani
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D017173:Caenorhabditis elegans / Q000235:genetics*; D016678:Genome / Q000187:drug effects / Q000528:radiation effects; D019076:Transgenes / Q000187:drug effects / Q000528:radiation effects; D014307:Trioxsalen / Q000494:pharmacology; D014466:Ultraviolet Rays
chemical_list: D014307:Trioxsalen
keywords: Caenorhabditis elegans; Genomic integration; Transgene; Trimethylpsoralen; Ultraviolet