title: Generation of a high-capacity hybrid vector: packaging of recombinant adenoassociated virus replicative intermediates in adenovirus capsids overcomes the limited cloning capacity of adenoassociated virus vectors.
abstract: Gene therapy aims to complement or, ideally, correct defective genes. The broad clinical application of this emerging technology requires the development of safe high-capacity gene delivery vehicles that combine efficient transduction of dividing as well as quiescent cells with sustained transgene expression. Here we present a new hybrid vector system that unites favorable attributes of adenoassociated virus (AAV) and adenovirus (Ad) vectors in a single particle. This was achieved by inclusion of Ad packaging elements in different sized recombinant AAV genomes. In the presence of AAV replicative functions and a recombinant helper Ad, AAV/Ad hybrid particles were generated via encapsidation of AAV-dependent replicative intermediates into Ad capsids. In stringent in vitro models based on transduction of proliferating cells we show that AAV/Ad hybrid vectors are superior to Ad vectors in establishing prolonged transgene expression and can be used to deliver DNA fragments of at least 27 kb.
authors.affiliation: Gene Therapy Section, Department of Molecular Cell Biology, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands. M.Goncalves@lumc.nl
authors.forename: M A
authors.initials: MA
authors.lastname: Gonçalves
authors.forename: M G
authors.initials: MG
authors.lastname: Pau
authors.forename: A A
authors.initials: AA
authors.lastname: de Vries
authors.forename: D
authors.initials: D
authors.lastname: Valerio
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics* / Q000502:physiology; D002213:Capsid / Q000235:genetics*; D002461:Cell Line, Transformed; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics* / Q000502:physiology; D016679:Genome, Viral; D006801:Humans; D011995:Recombination, Genetic; D019076:Transgenes; D014771:Virion; D019065:Virus Assembly / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication / Q000235:genetics / Q000502:physiology*