title: Deletion of genes from the mouse genome using Cre/loxP technology.
abstract: The steps required to delete genes from the mouse genome are illustrated by showing how a cluster of three flavin-containing monooxygenase (Fmo) genes (Fmol, Fmo2, and Fmo4) were deleted from mouse chromosome 1. Such large deletions are accomplished using loxP/Cre recombinase technology. Genomic clones corresponding to the genes to be deleted are first isolated, and then appropriate genomic fragments are cloned into vectors containing a loxP site. This produces targeting vectors, which are electroporated into mouse embryonic stem (ES) cells to allow a homologous recombination event to take place between the mouse genomic fragment, present within the vector, and the homologous sequences in the ES cell genome. Screening of ES cells for recombinants in which loxP sites have been inserted on either side of the gene cluster to be deleted is described. Recombination by Cre recombinase to produce ES cell lines carrying the deletion on chromosome 1 is also described.
authors.affiliation: Department of Biochemistry and Molecular Biology, University College London, UK.
authors.forename: Diana
authors.initials: D
authors.lastname: Hernandez
authors.forename: Pritpal
authors.initials: P
authors.lastname: Chandan
authors.forename: Azara
authors.initials: A
authors.lastname: Janmohamed
authors.forename: lan R
authors.initials: lR
authors.lastname: Phillips
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Shephard
mesh_terms: D000818:Animals; D017353:Gene Deletion; D016678:Genome; D019426:Integrases / Q000378:metabolism*; D007621:Karyotyping; D051379:Mice
chemical_list: C045073:Cre recombinase; D019426:Integrases