title: Production of a heterozygous mutant cell line by homologous recombination (single knockout).
abstract: Gene targeting by homologous recombination is a powerful and widely used technique for introduction of specific gene mutations (frequently a gene inactivation) in transgenic animals. The basic method detailed in this unit uses sequences homologous to the endogenous gene flanking the mutation. While methods using bacterial artificial chromosomes (BACs) and recombineering may be used, in most cases simpler bacterial plasmid clones with several kb of homology are sufficient. This protocol details the strategic factors in designing the constructs for selection and screening for homologous recombination.
authors.affiliation: University of Michigan Medical School, Ann Arbor, Michigan, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Mortensen
mesh_terms: D002460:Cell Line; D006579:Heterozygote; D009154:Mutation; D011995:Recombination, Genetic