Generation of medaka gene knockout models by target-selected mutagenesis.
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.
Taniguchi
Yoshihito
Y
Department of Radiation Genetics, CREST, Japan Science and Technology Laboratory, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan. .
Takeda
Shunichi
S
Furutani-Seiki
Makoto
M
Kamei
Yasuhiro
Y
Todo
Takeshi
T
Sasado
Takao
T
Deguchi
Tomonori
T
Kondoh
Hisato
H
Mudde
Josine
J
Yamazoe
Mitsuyoshi
M
Hidaka
Masayuki
M
Mitani
Hiroshi
H
Toyoda
Atsushi
A
Sakaki
Yoshiyuki
Y
Plasterk
Ronald H A
RH
Cuppen
Edwin
E
D000251:Adenosine Triphosphatases / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D004265:DNA Helicases / Q000235:genetics; D017931:DNA Primers; D005038:Ethylnitrosourea / Q000008:administration & dosage; D005260:Female; D016158:Genes, p53; D008297:Male; D008957:Models, Genetic; D016297:Mutagenesis, Site-Directed; D009990:Oryzias / Q000235:genetics*; D017354:Point Mutation; D053484:RecQ Helicases; D044767:Ubiquitin-Protein Ligases / Q000235:genetics
D017931:DNA Primers; D044767:Ubiquitin-Protein Ligases; C111567:parkin protein; D000251:Adenosine Triphosphatases; C097151:Bloom syndrome protein; D004265:DNA Helicases; D053484:RecQ Helicases; D005038:Ethylnitrosourea