| siRNA Screening for Genes Involved in HSV-1 Replication
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Author:
Date:
2014-08-20
[Abstract] Small interfering RNAs (siRNAs) are small (typically 18-24 nucleotides) RNA molecules capable of silencing gene expression post-transcriptionally and as such, they provide a simple method by which the role of individual genes in complex cellular systems can be easily assessed. As siRNAs are easy to use experimentally, and can be designed to target any gene (including pathogens), their use is perfectly suited to and easily adapted to high-throughput genome-wide screening methodologies and a range of phenotypic assays. Here we describe the use of a large siRNA library (>8,000 genes targeted individually) to screen for and identify host factors functionally involved in the replication of a human herpesvirus (Herpes simplex virus type 1; HSV-1) (Griffiths et al., 2013; Griffiths, ...
[摘要] 小干扰RNA（siRNA）是能够沉默转录后基因表达的小（通常为18-24个核苷酸）RNA分子，因此，它们提供了一种简单的方法，通过其可以容易地评估单个基因在复杂细胞系统中的作用。 由于siRNA易于实验使用，并且可以设计为靶向任何基因（包括病原体），它们的使用完全适合于并且容易地适应于高通量全基因组筛选方法学和一系列表型测定。 这里我们描述了使用大的siRNA文库（> 8,000个基因单独靶向）来筛选和鉴定功能上涉及人疱疹病毒（1型单纯疱疹病毒; HSV-1）（Griffiths）的复制的宿主因子， et al。，2013; Griffiths，2013）。
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| VZV Replication Assays
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Author:
Date:
2014-08-20
[Abstract] Varicella zoster virus (VZV) is a human herpesvirus which causes Varicella (chickenpox) upon primary infection and Zoster (shingles) following reactivation from latency (von Bokay, 1909). Whilst VZV is extensively studied, inherent features of VZV replication, such as cell-association of virus particles during in vitro culture and a restricted host range (limited to humans and some other primates) mean the cellular and viral mechanisms underlying VZV reactivation and pathogenesis remain largely uncharacterised. Much remains to be learnt about VZV, interactions with its host, and the development of disease. This protocol describes a basic VZV replication assay using a recombinant VZV-GFP reporter virus. As VZV is highly cell-associated in tissue culture, the reporter virus ...
[摘要] 水痘带状疱疹病毒（VZV）是人疱疹病毒，其在初次感染时引起水痘（水痘），在从潜伏期再活化后引起带状疱疹（带状疱疹）（von Bokay，1909）。虽然VZV被广泛研究，VZV复制的固有特征，例如在体外培养期间病毒颗粒的细胞缔合和受限的宿主范围（限于人和一些其它灵长类动物）意味着细胞和病毒基本的VZV再激活和发病机制仍然很大程度上没有表征。关于VZV，与其宿主的相互作用和疾病的发展仍然有很多知识。该协议描述了使用重组VZV-GFP报道病毒的基本VZV复制测定。由于VZV在组织培养中是高度细胞相关的，这里描述的报道病毒接种物是感染细胞的制备物。这种报道病毒感染的细胞系可以与siRNA基因缺失或cDNA过表达转染方案组合使用，以确定单个细胞基因对病毒复制的影响。
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