Griffiths Samantha J, Haas Jürgen
Division of Pathway and Infection Medicine, University of Edinburgh, Edinburgh, UK.
Bio Protoc. 2014 Aug 20;4(16). doi: 10.21769/BioProtoc.1209.
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 , 2013; Griffiths, 2013).
小干扰RNA(siRNA)是小的(通常为18 - 24个核苷酸)RNA分子,能够在转录后使基因表达沉默,因此,它们提供了一种简单的方法,通过该方法可以轻松评估复杂细胞系统中单个基因的作用。由于siRNA在实验中易于使用,并且可以设计成靶向任何基因(包括病原体),它们的使用非常适合并易于适应高通量全基因组筛选方法和一系列表型分析。在这里,我们描述了使用一个大型siRNA文库(单独靶向>8000个基因)来筛选和鉴定在人类疱疹病毒(单纯疱疹病毒1型;HSV - 1)复制中起功能作用的宿主因子(格里菲思,2013年;格里菲思,2013年)。