title: Targeted integration in rat and mouse embryos with zinc-finger nucleases.
abstract: Gene targeting is indispensible for reverse genetics and the generation of animal models of disease. The mouse has become the most commonly used animal model system owing to the success of embryonic stem cell-based targeting technology, whereas other mammalian species lack convenient tools for genome modification. Recently, microinjection of engineered zinc-finger nucleases (ZFNs) in embryos was used to generate gene knockouts in the rat and the mouse by introducing nonhomologous end joining (NHEJ)-mediated deletions or insertions at the target site. Here we use ZFN technology in embryos to introduce sequence-specific modifications (knock-ins) by means of homologous recombination in Sprague Dawley and Long-Evans hooded rats and FVB mice. This approach enables precise genome engineering to generate modifications such as point mutations, accurate insertions and deletions, and conditional knockouts and knock-ins. The same strategy can potentially be applied to many other species for which genetic engineering tools are needed.
authors.affiliation: Sigma Advanced Genetic Engineering Labs, Sigma-Aldrich Biotechnology, St. Louis, Missouri, USA. xiaoxia.cui@sial.com
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Cui
authors.forename: Diana
authors.initials: D
authors.lastname: Ji
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Fisher
authors.forename: Yumei
authors.initials: Y
authors.lastname: Wu
authors.forename: David M
authors.initials: DM
authors.lastname: Briner
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Weinstein
mesh_terms: D000818:Animals; D001483:Base Sequence; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics*; D004622:Embryo, Mammalian / Q000378:metabolism*; D053595:Embryonic Stem Cells; D055786:Gene Knockout Techniques / Q000379:methods; D018390:Gene Targeting; D005818:Genetic Engineering / Q000379:methods*; D051379:Mice; D008845:Microinjections; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011995:Recombination, Genetic; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D011993:Recombinant Fusion Proteins; D015252:Deoxyribonucleases, Type II Site-Specific