Targeted disruption of exogenous EGFP gene in medaka using zinc-finger nucleases

Satoshi Ansai, Hiroshi Ochiai, Yuta Kanie, Yasuhiro Kamei, Yuki Gou, Takeshi Kitano, Takashi Yamamoto, Masato Kinoshita

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Zinc-finger nucleases (ZFNs) are artificial enzymes that create site-specific double-strand breaks and thereby induce targeted genome editing. Here, we demonstrated successful gene disruption in somatic and germ cells of medaka (Oryzias latipes) using ZFN to target exogenous EGFP genes. Embryos that were injected with an RNA sequence pair coding for ZFNs showed mosaic loss of green fluorescent protein fluorescence in skeletal muscle. A number of mutations that included both deletions and insertions were identified within the ZFN target site in each embryo, whereas no mutations were found at the non-targeted sites. In addition, ZFN-induced mutations were introduced in germ cells and efficiently transmitted to the next generation. The mutation frequency varied (6-100%) in the germ cells from each founder, and a founder carried more than two types of mutation in germ cells. Our results have introduced the possibility of targeted gene disruption and reverse genetics in medaka.

Original languageEnglish
Pages (from-to)546-556
Number of pages11
JournalDevelopment Growth and Differentiation
Volume54
Issue number5
DOIs
Publication statusPublished - 2012 Jun 1
Externally publishedYes

Keywords

  • Gene knock-out
  • Germ-line transmission
  • Green fluorescent protein
  • Medaka
  • Zinc-finger nucleases

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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