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Generation of medaka gene knockout models by target-selected mutagenesis |
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| Authors: | 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 HA Plasterk Edwin Cuppen |
| Institution: | Department of Radiation Genetics, CREST, Japan Science and Technology Laboratory, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan. . |
| Abstract: | 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. |
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