1. Generation of medaka gene knockout models by target-selected mutagenesis
- Author
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Yoshihito, Taniguchi, Shunichi, Takeda, Makoto, Furutani-Seiki, Yasuhiro, Kamei, Takeshi, Todo, Takao, Sasado, Tomonori, Deguchi, Hisato, Kondoh, Josine, Mudde, Mitsuyoshi, Yamazoe, Masayuki, Hidaka, Hiroshi, Mitani, Atsushi, Toyoda, Yoshiyuki, Sakaki, Ronald H A, Plasterk, and Edwin, Cuppen
- Subjects
Adenosine Triphosphatases ,Male ,animal structures ,Base Sequence ,Models, Genetic ,RecQ Helicases ,Ubiquitin-Protein Ligases ,fungi ,DNA Helicases ,Oryzias ,Method ,Genes, p53 ,Animals, Genetically Modified ,Ethylnitrosourea ,embryonic structures ,Mutagenesis, Site-Directed ,Animals ,Point Mutation ,Female ,human activities ,reproductive and urinary physiology ,DNA Primers - Abstract
A reverse genetics approach for the routine generation of medaka (Oryzias latipes) gene knockouts is described and applied to create a cryopreserved resource containing knockouts for most medaka genes., We have established a reverse genetics approach for the routine generation of medaka (Oryzias latipes) gene knockouts. A cryopreserved library of N-ethyl-N-nitrosourea (ENU) mutagenized fish was screened by high-throughput resequencing for induced point mutations. Nonsense and splice site mutations were retrieved for the Blm, Sirt1, Parkin and p53 genes and functional characterization of p53 mutants indicated a complete knockout of p53 function. The current cryopreserved resource is expected to contain knockouts for most medaka genes.
- Published
- 2006