Agarwalla Pankaj K, Aghi Manish K
Harvard Medical School, Boston, MA, USA.
Methods Mol Biol. 2012;797:1-19. doi: 10.1007/978-1-61779-340-0_1.
Herpes simplex virus-1 (HSV-1) is an enveloped, double-stranded DNA virus that has been used with modification as an oncolytic virus against a number of tumor types. Modifications that make HSV-1 replication--conditional, i.e., selectively divide in replicating cells make it fulfill a prerequisite criteria for oncolytic viruses. Other appealing features of HSV-1 as an oncolytic virus include its large, modifiable genome; its sensitivity to antiviral agents, such as ganciclovir; and its lack of host cell integration. Here, we review the methods of HSV-1 engineering, through traditional recombination techniques as well as through bacterial artificial chromosome (BAC) technology. We then describe protocols for titering, amplification, and purification of engineered HSV-1-derived oncolytic viruses.
单纯疱疹病毒1型（HSV-1）是一种有包膜的双链DNA病毒，已通过修饰用作针对多种肿瘤类型的溶瘤病毒。使HSV-1复制具有条件性，即在复制细胞中选择性分裂的修饰使其满足溶瘤病毒的一个先决标准。HSV-1作为溶瘤病毒的其他吸引人的特性包括其庞大的、可修饰的基因组；其对更昔洛韦等抗病毒药物的敏感性；以及其缺乏宿主细胞整合。在这里，我们回顾了通过传统重组技术以及细菌人工染色体（BAC）技术对HSV-1进行工程改造的方法。然后，我们描述了工程化HSV-1衍生的溶瘤病毒的滴度测定、扩增和纯化方案。
