Site-specific modification of genome with cell-permeable Cre fusion protein in preimplantation mouse embryo

Kyoungmi Kim, Hwain Kim, Daekee Lee

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Site-specific recombination (SSR) by Cre recombinase and its target sequence, loxP, is a valuable tool in genetic analysis of gene function. Recently, several studies reported successful application of Cre fusion protein containing protein transduction peptide for inducing gene modification in various mammalian cells including ES cell as well as in the whole animal. In this study, we show that a short incubation of preimplantation mouse embryos with purified cell-permeable Cre fusion protein results in efficient SSR. X-Gal staining of preimplantation embryos, heterozygous for Gtrosa26tm1Sor, revealed that treatment of 1-cell or 2-cell embryos with 3 μM of Cre fusion protein for 2 h leads to Cre-mediated excision in 70-85% of embryos. We have examined the effect of the concentration of the Cre fusion protein and the duration of the treatment on embryonic development, established a condition for full term development and survival to adulthood, and demonstrated the germ line transmission of excised Gtrosa26 allele. Potential applications and advantages of the highly efficient technique described here are discussed.

Original languageEnglish
Pages (from-to)122-126
Number of pages5
JournalBiochemical and biophysical research communications
Volume388
Issue number1
DOIs
Publication statusPublished - 2009 Oct 9
Externally publishedYes

Keywords

  • Cre recombinase
  • Embryonic development
  • Germ line transmission
  • Preimplantation embryo
  • Protein transduction
  • ROSA26
  • Site-specific recombination

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Site-specific modification of genome with cell-permeable Cre fusion protein in preimplantation mouse embryo'. Together they form a unique fingerprint.

Cite this