title: HIV vector production mediated by Rev protein transduction.
abstract: HIV-1-based vectors are promising tools for gene therapy because of their ability to integrate into nondividing cells. Their safety in clinical applications remains a major concern. Recombination events occurring among plasmid constructs during vector production could potentially lead to the generation of replication-competent viruses. The safety of HIV-1-based vectors can be improved by removing all regions of the viral genome that are not absolutely required for vector production or function. In this study, we demonstrate that the HIV-1 rev gene is dispensable for the production of HIV-1-based vectors if the vector-producing cells are supplied with purified Rev protein. We compared the efficiency of vector production among Rev, TAT-Rev (Rev fused to the protein transduction domain of the HIV TAT protein), and Rev/Pep-1 (Rev complexed with the carrier peptide Pep-1). Our results showed that 293T cells efficiently internalized TAT-Rev and Rev/Pep-1 and high-titer vector preparations were obtained with this approach. Vectors generated by such an approach showed little difference in their efficiencies of transduction of established cell lines and primary cells compared with vectors generated by standard plasmid cotransfection. Eliminating the requirement for the HIV-1 rev gene during vector production should improve the safety of applying HIV vectors in human clinical trials.
authors.affiliation: Department of Virology, Beckman Research Institute, City of Hope, 1500 E. Duarte Road, Duarte, California 91010, USA.
authors.forename: Claudia M
authors.initials: CM
authors.lastname: Kowolik
authors.forename: Priscilla
authors.initials: P
authors.lastname: Yam
authors.forename: Ying
authors.initials: Y
authors.lastname: Yu
authors.forename: Jiing-Kuan
authors.initials: JK
authors.lastname: Yee
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D015967:Gene Expression Regulation, Viral; D015694:Gene Products, rev / Q000235:genetics / Q000378:metabolism*; D015756:Genes, tat / Q000235:genetics; D005822:Genetic Vectors / Q000096:biosynthesis* / Q000235:genetics; D006678:HIV / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000235:genetics; D054321:rev Gene Products, Human Immunodeficiency Virus
chemical_list: D002352:Carrier Proteins; D015694:Gene Products, rev; D054321:rev Gene Products, Human Immunodeficiency Virus; D001616:beta-Galactosidase