title: Site-specific genomic integration in mammalian cells mediated by phage phiC31 integrase.
abstract: We previously established that the phage phiC31 integrase, a site-specific recombinase, mediates efficient integration in the human cell environment at attB and attP phage attachment sites on extrachromosomal vectors. We show here that phage attP sites inserted at various locations in human and mouse chromosomes serve as efficient targets for precise site-specific integration. Moreover, we characterize native "pseudo" attP sites in the human and mouse genomes that also mediate efficient integrase-mediated integration. These sites have partial sequence identity to attP. Such sites form naturally occurring targets for integration. This phage integrase-mediated reaction represents an effective site-specific integration system for higher cells and may be of value in gene therapy and other chromosome engineering strategies.
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305-5120.
authors.forename: B
authors.initials: B
authors.lastname: Thyagarajan
authors.forename: E C
authors.initials: EC
authors.lastname: Olivares
authors.forename: R P
authors.initials: RP
authors.lastname: Hollis
authors.forename: D S
authors.initials: DS
authors.lastname: Ginsburg
authors.forename: M P
authors.initials: MP
authors.lastname: Calos
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001435:Bacteriophages / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D016678:Genome; D006801:Humans; D019426:Integrases / Q000502:physiology*; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D012641:Selection, Genetic; D012689:Sequence Homology, Nucleic Acid; D016662:Virus Integration / Q000235:genetics* / Q000502:physiology*
chemical_list: D017931:DNA Primers; D004247:DNA; D008156:Luciferases; D019426:Integrases