Measuring recombination proficiency in mouse embryonic stem cells

Methods Mol Biol. 2005;291:373-84. doi: 10.1385/1-59259-840-4:373.

Abstract

A method is presented to measure homologous recombination in mouse embryonic stem cells by both gene targeting and short-tract gene conversion of a double-strand break. A fluorescence-based reporter is first gene targeted to the Hprt locus in a quantifiable way. A homing endonuclease expression vector is then introduced to generate a double-strand break, the repair of which is also quantifiable.

MeSH terms

  • Animals
  • DNA / analysis
  • DNA Damage / genetics
  • DNA Repair / genetics
  • Embryo, Mammalian / cytology*
  • Gene Conversion / genetics*
  • Gene Targeting*
  • Hypoxanthine Phosphoribosyltransferase / genetics
  • Mice
  • Recombination, Genetic / genetics*
  • Stem Cells / chemistry*

Substances

  • DNA
  • Hypoxanthine Phosphoribosyltransferase