title: Targeted homologous recombination at the endogenous adenine phosphoribosyltransferase locus in Chinese hamster cells.
abstract: We have developed a system that permits analysis of targeted homologous recombination at an endogenous, chromosomal gene locus in cultured mammalian cells. Using a hemizygous, adenine phosphoribosyltransferase (APRT)-deficient, Chinese hamster ovary (CHO) cell mutant as a transfection recipient, we have demonstrated correction of a nonrevertible deletion mutation by targeted homologous recombination. Transfection with a plasmid carrying a fragment of the APRT gene yielded APRT+ recombinants at a frequency of approximately 4.1 x 10(-7). The ratio of targeted recombination to nontargeted integrations of plasmid sequences was approximately 1:4000. Analysis of 31 independent APRT+ recombinants revealed conversions of the endogenous APRT gene, targeted integration at the APRT locus, and a third class of events in which the plasmid donor APRT fragment was converted to a full-length, functional gene.
authors.affiliation: University of Texas M. D. Anderson Cancer Center, Science Park-Research Division, Smithville 78957.
authors.forename: G M
authors.initials: GM
authors.lastname: Adair
authors.forename: R S
authors.initials: RS
authors.lastname: Nairn
authors.forename: J H
authors.initials: JH
authors.lastname: Wilson
authors.forename: M M
authors.initials: MM
authors.lastname: Seidman
authors.forename: K A
authors.initials: KA
authors.lastname: Brotherman
authors.forename: C
authors.initials: C
authors.lastname: MacKinnon
authors.forename: J B
authors.initials: JB
authors.lastname: Scheerer
mesh_terms: D000228:Adenine Phosphoribosyltransferase / Q000172:deficiency / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D002874:Chromosome Mapping; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D005796:Genes; D005822:Genetic Vectors; D009693:Nucleic Acid Hybridization; D010053:Ovary; D010430:Pentosyltransferases / Q000235:genetics*; D011995:Recombination, Genetic; D014162:Transfection
chemical_list: D010430:Pentosyltransferases; D000228:Adenine Phosphoribosyltransferase