Improved HSV-1 amplicon packaging using virion host shutoff mutants lacking mRNAse activity.
Improved HSV-1 amplicon packaging using virion host shutoff mutants lacking mRNAse activity.
复制标题
使用缺乏 mRNAse 活性的病毒体宿主关闭突变体改进了 HSV-1 扩增子包装。
DOI：
10.1002/jgm.972
复制
发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Federoff,HJ
中科院分区：
文献类型：
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作者：
Halterman,MW;Giuliano,RE;Bowers,WJ;Federoff,HJ
鉴于其慷慨的转基因能力和固有的亲神经性，单纯疱疹病毒（HSV-1）为基础的病毒载体是有前途的工具，基因输送到中枢神经系统。尽管它们在临床前广泛使用，但在人类中使用疱疹病毒载体时，载体毒性仍然是一个问题。一个潜在的毒性来源来自被膜相关病毒体宿主关闭蛋白（vhs），其通过非特异性mRNA酶活性诱导宿主细胞中的翻译停滞。在当前的研究中，我们使用了一系列在UL 41开放阅读框架中含有缺失的HSV-1病毒来研究：（1）扩增子包装中完整vhs功能的要求和（2）vhs是否影响解离皮层神经元的转导后存活。我们的结果表明，虽然扩增子产量降低了一个数量级，但UL 41缺失与载体毒性降低有关。此外，使用mRNAse失活点突变体部分重建vhs功能可改善扩增子滴度，而不会产生野生型对照所观察到的毒性。这些发现为改善HSV病毒载体的滴度和毒性特征提供了一种新方法。版权所有© 2006约翰威利父子有限公司。
Given their generous transgene capacity and inherent neurotropism, herpes simplex virus (HSV‐1)‐based viral vectors are promising tools for gene delivery to the central nervous system. Despite their widespread pre‐clinical use, vector toxicity remains a concern with regard to the use of herpes vectors in humans. One potential source of toxicity stems from the tegument‐associated virion host shutoff protein (vhs), which induces translational arrest in the host cell through non‐specific mRNAse activity. In the current study we utilized a series of HSV‐1 viruses containing a deletion in the UL41 open reading frame to investigate: (1) the requirement of intact vhs function in amplicon packaging and (2) whether vhs influences the post‐transduction survival of dissociated cortical neurons. Our results demonstrate that while amplicon yield was reduced an order of magnitude, UL41 deletion was associated with reduced vector toxicity. Furthermore, partial reconstitution of vhs function using mRNAse‐inactive point mutants improved amplicon titers without imparting the toxicity observed with wild‐type controls. These findings offer a novel approach to improving the titer and toxicity profiles of HSV‐based viral vectors. Copyright © 2006 John Wiley & Sons, Ltd.