Site-specific integration mediated by a hybrid adenovirus/adeno-associated virus vector
| Autor: | Riccardo Cortese, Alessandra Recchia, S. Colloca, Fabio Palombo, Frank L. Graham, Gennaro Ciliberto, N. La Monica, Carlo Toniatti, Luisa Pieroni, R. J. Parks, S. Lamartina |
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| Rok vydání: | 1999 |
| Předmět: |
site specific integration
viruses Genetic enhancement Transgene Genetic Vectors Molecular Sequence Data Biology medicine.disease_cause Virus Adenoviridae adenovirus AAV Transduction (genetics) medicine Humans DNA Integration Adeno-associated virus Gene In Situ Hybridization Multidisciplinary Base Sequence DNA Helicases Gene Transfer Techniques Dependovirus Biological Sciences Virology DNA-Binding Proteins Trans-Activators Reassortant Viruses HeLa Cells |
| Zdroj: | Proceedings of the National Academy of Sciences. 96:2615-2620 |
| ISSN: | 1091-6490 0027-8424 |
| Popis: | Adenovirus (Ad) and adeno-associated virus (AAV) have attractive and complementary properties that can be exploited for gene transfer purposes. Ad vectors are probably the most efficient vehicles to deliver foreign genes both in vitro and in vivo . AAV exhibits the unique ability to establish latency by efficiently integrating at a specific locus of human chromosome 19 (AAVS1). Two viral elements are necessary for the integration at AAVS1: Rep68/78 and the inverted terminal repeats (AAV-ITRs). In this study, we report the development of two helper-dependent adenoviral (HD) vectors, one carrying the Rep78 gene, the other an AAV-ITR-flanked transgene. Although Rep proteins have been demonstrated to interfere with Ad replication, HD Rep78 vector was successfully amplified on serial passages in 293CRE4 cells with a yield of 50–100 transducing units per cell. DNA integration at the AAVS1 site also was demonstrated in hepatoma cells coinfected with the HD-expressing Rep78 and with the second HD vector carrying a transgene flanked by AAV-ITRs. The high transduction efficiency, large cloning capacity, and high titer of the HD, combined with the site-specific integration machinery provided by AAV-derived components, make the Ad/AAV hybrid viruses a promising vehicle for gene therapy. |
| Databáze: | OpenAIRE |
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