title: Efficient method to generate single-copy transgenic mice by site-specific integration in embryonic stem cells.
abstract: Transgenic and gene-targeted mutant mice provide powerful tools for analysis of the cellular processes involved in early development and in the pathogenesis of many diseases. Here we describe a transgene integration strategy mediated by site-specific recombination that allows establishment of multiple embryonic stem (ES) cell lines carrying tetracycline-inducible genes targeted to a specific locus to assure predictable temporal and spatial expression in ES cells and mice. Using homologous recombination we inserted an frt homing site into which tetracycline-inducible transgenes can be integrated efficiently in the presence of FLPe recombinase. This strategy and the vectors described here are generally applicable to any locus in ES cells and should allow for the rapid production of mice with transgenes efficiently targeted to a defined site.
authors.affiliation: Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA.
authors.forename: Caroline
authors.initials: C
authors.lastname: Beard
authors.forename: Konrad
authors.initials: K
authors.lastname: Hochedlinger
authors.forename: Kathrin
authors.initials: K
authors.lastname: Plath
authors.forename: Anton
authors.initials: A
authors.lastname: Wutz
authors.forename: Rudolf
authors.initials: R
authors.lastname: Jaenisch
mesh_terms: D000483:Alleles; D000818:Animals; D024042:Collagen Type I / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology*; D004815:Epidermal Growth Factor / Q000235:genetics; D005434:Flow Cytometry; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D011995:Recombination, Genetic; D013234:Stem Cells / Q000378:metabolism*; D013752:Tetracycline / Q000494:pharmacology
chemical_list: D024042:Collagen Type I; D004815:Epidermal Growth Factor; D013752:Tetracycline