Generation of medaka gene knockout models by target-selected mutagenesis

Genome Biol. 2006;7(12):R116. doi: 10.1186/gb-2006-7-12-r116.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / genetics
  • Animals
  • Animals, Genetically Modified*
  • Base Sequence
  • DNA Helicases / genetics
  • DNA Primers
  • Ethylnitrosourea / administration & dosage
  • Female
  • Genes, p53
  • Male
  • Models, Genetic*
  • Mutagenesis, Site-Directed
  • Oryzias / genetics*
  • Point Mutation
  • RecQ Helicases
  • Ubiquitin-Protein Ligases / genetics

Substances

  • DNA Primers
  • Ubiquitin-Protein Ligases
  • parkin protein
  • Adenosine Triphosphatases
  • Bloom syndrome protein
  • DNA Helicases
  • RecQ Helicases
  • Ethylnitrosourea