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

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• 作者：Stuart Macnab ; Robert White ; Julian Hiscox ; Adrian Whitehouse
• 关键词：Gene therapy ; Herpesvirus saimiri ; Amplicon ; Episome
• 刊名：Journal of Biotechnology
• 出版年：2008
• 出版时间：30 April 2008
• 年：2008
• 卷：134
• 期：3-4
• 页码：287-296
• 全文大小：1397 K

文摘

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, OMKHVSΔBam, 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), OMKHVSΔBam 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.