Production of an infectious Herpesvirus saimiri-based episomally maintained amplicon system

J Biotechnol. 2008 Apr 30;134(3-4):287-96. doi: 10.1016/j.jbiotec.2008.01.012. Epub 2008 Jan 25.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Transformed
  • Cell Line, Tumor
  • Chromosomes, Artificial, Bacterial / metabolism
  • Defective Viruses / genetics*
  • Defective Viruses / pathogenicity
  • Female
  • Gene Transfer Techniques
  • Genome, Viral*
  • Helper Viruses / genetics
  • Herpesvirus 2, Saimiriine / genetics*
  • Herpesvirus 2, Saimiriine / pathogenicity
  • Recombination, Genetic
  • Transgenes / physiology
  • Virus Assembly / genetics