Production of an infectious Herpesvirus saimiri-based episomally maintained amplicon system.
Viral vectors have a number of obstacles to overcome for effective gene therapy, including immune stimulation, packaging potential and cell tropism. Herpesvirus saimiri (HVS) has many favourable traits including, a large packaging capability, wide cell tropism, and the ability to episomally persist as an artificial chromosome. To further develop HVS as a gene therapy vector we aim to produce a safe disabled HVS-based recombinant viral system for gene therapy applications. An HVS recombinant viral amplicon was constructed with a transgene packaging potential of 50 kb. The recombinant HVS genome was shown to be replication disabled and used to generate a stable cell line, OMKHVS Delta Bam, in which the modified genome persists as a non-integrated episome. To assess whether the modified genome could be packaged into a virus-like particle (HVSampVLP), OMKHVS Delta Bam was infected with replication competent virus or transfected with a defective helper virus. The resultant HVSampVLPs were able to infect SW480 tumour cells, delivering the recombinant disabled genome, which persisted as a non-integrated episome in the dividing cell population. This study forms the basis of a replication disabled HVS amplicon system for use in gene therapy applications.
Macnab
Stuart
S
Institute of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, United Kingdom.
White
Robert
R
Hiscox
Julian
J
Whitehouse
Adrian
A
D000818:Animals; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D022202:Chromosomes, Artificial, Bacterial / Q000378:metabolism; D003673:Defective Viruses / Q000235:genetics* / Q000472:pathogenicity; D005260:Female; D018014:Gene Transfer Techniques; D016679:Genome, Viral; D006378:Helper Viruses / Q000235:genetics; D006567:Herpesvirus 2, Saimiriine / Q000235:genetics* / Q000472:pathogenicity; D011995:Recombination, Genetic; D019076:Transgenes / Q000502:physiology; D019065:Virus Assembly / Q000235:genetics