title: Generation of medaka gene knockout models by target-selected mutagenesis.
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.
authors.affiliation: Department of Radiation Genetics, CREST, Japan Science and Technology Laboratory, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan. .
authors.forename: Yoshihito
authors.initials: Y
authors.lastname: Taniguchi
authors.forename: Shunichi
authors.initials: S
authors.lastname: Takeda
authors.forename: Makoto
authors.initials: M
authors.lastname: Furutani-Seiki
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kamei
authors.forename: Takeshi
authors.initials: T
authors.lastname: Todo
authors.forename: Takao
authors.initials: T
authors.lastname: Sasado
authors.forename: Tomonori
authors.initials: T
authors.lastname: Deguchi
authors.forename: Hisato
authors.initials: H
authors.lastname: Kondoh
authors.forename: Josine
authors.initials: J
authors.lastname: Mudde
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Yamazoe
authors.forename: Masayuki
authors.initials: M
authors.lastname: Hidaka
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Mitani
authors.forename: Atsushi
authors.initials: A
authors.lastname: Toyoda
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Sakaki
authors.forename: Ronald H A
authors.initials: RH
authors.lastname: Plasterk
authors.forename: Edwin
authors.initials: E
authors.lastname: Cuppen
mesh_terms: 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
chemical_list: 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