Measuring recombination proficiency in mouse embryonic stem cells.
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.
Pierce
Andrew J
AJ
Markey Cancer Center, University of Kentucky, Lexington, USA.
Jasin
Maria
M
D000818:Animals; D004247:DNA / Q000032:analysis; D004249:DNA Damage / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology*; D005785:Gene Conversion / Q000235:genetics*; D018390:Gene Targeting; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D051379:Mice; D011995:Recombination, Genetic / Q000235:genetics*; D013234:Stem Cells / Q000737:chemistry*
D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase