title: Phage integrases for the construction and manipulation of transgenic mammals.
abstract: Phage integrases catalyze site-specific, unidirectional recombination between two short att recognition sites. Recombination results in integration when the att sites are present on two different DNA molecules and deletion or inversion when the att sites are on the same molecule. Here we demonstrate the ability of the phiC31 integrase to integrate DNA into endogenous sequences in the mouse genome following microinjection of donor plasmid and integrase mRNA into mouse single-cell embryos. Transgenic early embryos and a mid-gestation mouse are reported. We also demonstrate the ability of the phiC31, R4, and TP901-1 phage integrases to recombine two introduced att sites on the same chromosome in human cells, resulting in deletion of the intervening material. We compare the frequencies of mammalian chromosomal deletion catalyzed by these three integrases in different chromosomal locations. The results reviewed here introduce these bacteriophage integrases as tools for site-specific modification of the genome for the creation and manipulation of transgenic mammals.
authors: Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305-5120, USA. RHollis@chla.usc.edu
authors: Roger P
authors: RP
authors: Hollis
authors: Stephanie M
authors: SM
authors: Stoll
authors: Christopher R
authors: CR
authors: Sclimenti
authors: Jennifer
authors: J
authors: Lin
authors: Yanru
authors: Y
authors: Chen-Tsai
authors: Michele P
authors: MP
authors: Calos
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics; D017103:Bacillus Phages / Q000201:enzymology*; D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D018390:Gene Targeting / Q000379:methods*; D016678:Genome; D006801:Humans; D019426:Integrases / Q000378:metabolism*; D008322:Mammals / Q000196:embryology / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D019076:Transgenes / Q000235:genetics*
chemical_list: D019426:Integrases