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 ...
[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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Dharmacon siRNA Transfection of HeLa Cells
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Author:
Date:
2012-02-20
[Abstract] Small Interfering RNA (siRNA) is a class of double-stranded RNAs of 20-25 nucleotides that play important roles in many biological processes (Hamilton and Baulcombe, 1999). siRNAs act by “neutralizing” the mRNA of the target protein, facilitating degradation of the mRNA and hence altering the biological effect of the protein (reviewed in Hannon and Rossi, 2004). siRNAs may also change the intracellular levels of regulatory RNAs. Use of siRNAs for manipulating the expression of genes of interest in biological research is commonly referred to as RNA interference or knockdown technique (Elbashir et ...
[摘要] 小干扰RNA (siRNA)是一类20~25nt大小的双链RNA,在生物体内起着重要的作用 [1]. 人工合成的siRNAs 已经成为一个有效的工具广泛的应用于调控基因转录和翻译,它可以作用于感兴趣的靶向蛋白使其mRNA失效(“敲除”),这一过程通常被称为RNA干扰[2].RNA干扰(敲除)是一个使靶向RNA降解的复杂过程[3 & refs therein]. 对同一靶向RNA,通过不同的公司可能设计的干扰小RNA不同,这里介绍一种用Dharmacon (Thermo)的 siGENOME SMARTpool试剂介导的siRNA转染的实验方法 。
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