A novel herpesvirus amplicon system for in vivo gene delivery.
For gene therapy approaches to succeed, improved vector systems are needed that combine a large carrying capacity with high transduction efficiency in vivo. Towards this goal, we have developed a novel herpes simplex virus (HSV) amplicon vector, pHE, which contains an HSV-1 replication origin (ori S) and packaging sequence that permit vector replication and packaging into HSV-1 capsids. The vector also contains the Epstein-Barr virus (EBV) unique latent replication origin (ori P) sequence and a modified EBNA-1 gene to allow the vector to be maintained as an episome in transfected E5 helper cells. This system allows for efficient packaging of high-titer vector since the E5 cells are first selected for the presence of the pHE vector before helper virus infection. The infectious pHE vector has efficient transgene expression in a variety of human cell lines in vitro. Stereotactic injection of pHE vector supernatant into the rat brain resulted in high, localized reporter gene expression. Finally, the pHE vector could carry a stable 21 kb DNA payload into HSV virions. This pHE vector system should have a broad range of gene transfer applications.
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Human Gene Therapy Research Institute, Iowa Health System, Des Moines 50309, USA.
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Jacobson
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D000818:Animals; D001345:Autoradiography; D015139:Blotting, Southern; D002490:Central Nervous System; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics*; D005784:Gene Amplification; D015870:Gene Expression; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human; D004854:Herpesvirus 4, Human; D006801:Humans; D010957:Plasmids; D051381:Rats; D019076:Transgenes; D014407:Tumor Cells, Cultured
D019309:Epstein-Barr Virus Nuclear Antigens