title: Measuring recombination proficiency in mouse embryonic stem cells.
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 (DSB). 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 DSB, the repair of which is also quantifiable.
authors.affiliation: Markey Cancer Center, University of Kentucky, Lexington, KY, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Pierce
authors.forename: Maria
authors.initials: M
authors.lastname: Jasin
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D053903:DNA Breaks, Double-Stranded; D053595:Embryonic Stem Cells / Q000502:physiology*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D007041:Hypoxanthine Phosphoribosyltransferase / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000235:genetics; D059767:Recombinational DNA Repair; D017422:Sequence Analysis, DNA; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins; D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase