Generation of medaka gene knockout models by target-selected mutagenesis

Yoshihito Taniguchi¹ , Shunichi Takeda¹ , Makoto Furutani-Seiki² , Yasuhiro Kamei³ , Takeshi Todo³ , Takao Sasado² , Tomonori Deguchi² , Hisato Kondoh² , Josine Mudde⁴ , Mitsuyoshi Yamazoe¹ , Masayuki Hidaka⁵ , Hiroshi Mitani⁵ , Atsushi Toyoda⁶ , Yoshiyuki Sakaki⁶ , Ronald HA Plasterk⁴ and Edwin Cuppen⁴

¹Department of Radiation Genetics, CREST, Japan Science and Technology Laboratory, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan

²Kondoh Differentiation Signaling Project, Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Corporation, Yoshida-kawaramachi, Sakyo-ku, Kyoto, 606-8305, Japan

³Department of Mutagenesis, Radiation Biology Center, Kyoto University, Yoshida Konoe, Sakyoku, Kyoto 606-8501, Japan

⁴Hubrecht Laboratory, Uppsalalaan, Utrecht, The Netherlands

⁵Department of Integrated Biosciences, The University of Tokyo, 5-1-5 Kashiwa-no-ha, Kashiwa, Chiba 277-8562, Japan

⁶The Institute of Physical and Chemical Research Genomic Sciences Center, RIKEN Yokohama Institute, 1-7-22 Suehiro, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan

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Genome Biology 2006, 7:R116doi:10.1186/gb-2006-7-12-r116


Published:	8 December 2006

Subject areas: Development, Genetics, Model organisms

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.