title: Site-specific integration of bacterial artificial chromosomes into human cells.
abstract: Gene therapy of inherited diseases requires long-term maintenance of the corrective transgene. Stable integration of the introduced DNA molecule into one of the host cell chromosomes is the simplest strategy for achieving this. However, genotoxicity resulting from random insertion of the transgene raises serious safety concerns that must be addressed if gene therapy is to enter the clinical mainstream. The following method makes use of the Rep integrase of adeno-associated virus to insert a transgene into the human AAVS1 site, a known "safe harbor" region within the human genome. This approach has the potential for application to novel gene therapy strategies for improved safety. In addition, with this method it is also possible to create cell lines carrying BAC transgenes in the AAVS1 site.
authors.affiliation: Cell and Gene Therapy Research Group, Murdoch Children's Research Institute, Royal Children's Hospital, Flemington Road, Parkville, 3052, Melbourne, VIC, Australia, brad.mccoll@mcri.edu.au.
authors.forename: Bradley
authors.initials: B
authors.lastname: McColl
authors.forename: Sara
authors.initials: S
authors.lastname: Howden
authors.forename: Jim
authors.initials: J
authors.lastname: Vadolas
mesh_terms: D001483:Base Sequence; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D016662:Virus Integration / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D012367:RNA, Viral; D014764:Viral Proteins; C068734:rep proteins, Adeno-associated virus 2; D019426:Integrases