Generation of medaka gene knockout models by target-selected mutagenesis.
Taniguchi Y,
Takeda S,
Furutani-Seiki M,
Kamei Y,
Todo T,
Sasado T,
Deguchi T,
Kondoh H,
Mudde J,
Yamazoe M,
Hidaka M,
Mitani H,
Toyoda A,
Sakaki Y,
Plasterk RH,
Cuppen E.
Department of Radiation Genetics, CREST, Japan Science and Technology Laboratory, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan. .
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.
PMID: 17156454 [PubMed - indexed for MEDLINE]
PMCID: PMC1794429