| Genome-Wide siRNA Screen for Anti-Cancer Drug Resistance in Adherent Cell Lines
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Author:
Date:
2015-05-20
[Abstract] The expression of genes is frequently manipulated in cell lines to study their cellular functions. The use of exogenous small Interfering RNAs (siRNAs) is a very efficient technique to temporarily downregulate the expression of genes of interest [reviewed by Hannon and Rossi (2004)]. A genome-wide siRNA library allows the user to study both the effect of each individual gene on a particular cell phenotype in a high throughput manner and also assess its phenotypic effect relative to all other genes targeted. Several factors that potentially influence the outcome of a screen need to be considered when performing a large siRNA screen (Jiang et al., 2011). Here we present a detailed protocol for a genome-wide screen to identify genes involved in anti-cancer drug resistance using the ...
[摘要] 基因的表达经常在细胞系中操作以研究它们的细胞功能。外源小干扰RNA（siRNA）的使用是临时下调感兴趣的基因的表达的非常有效的技术[由Hannon和Rossi（2004）综述]。全基因组siRNA文库允许用户以高通量方式研究每个个体基因对特定细胞表型的作用，并且还评估其相对于所有其他靶向基因的表型效应。当进行大siRNA筛选时，需要考虑潜在地影响筛选结果的几个因素（Jiang等人，2011）。在这里，我们提出了一个详细的协议，使用人类siGENOME库从Dharmacon的基因组范围屏幕识别参与抗癌耐药性的基因。在该方案中，我们关注对表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKI）厄洛替尼在对EGFR-TKIs敏感的肺癌细胞系PC9中的治疗的抗性（de Bruin等。，2014）。该方案可用于其他细胞系和其他药物治疗，我们在下面的注释中扩展。
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| 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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| 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 al., 2001). Synthetic siRNAs are an emerging tool that are now widely used in these studies. A variety of algorithms are employed by different companies for the design of siRNA products, ...
[摘要] 小干扰RNA（siRNA）是一类20-25个核苷酸的双链RNA，在许多生物过程中起重要作用（Hamilton和Baulcombe，1999）。 siRNA通过"中和"靶蛋白的mRNA起作用，促进mRNA的降解，从而改变蛋白的生物效应（综述于Hannon和Rossi，2004）。 siRNA也可以改变调节RNA的细胞内水平。 在生物研究中使用siRNA操纵目的基因的表达通常被称为RNA干扰或敲除技术（Elbashir等人，2001）。 合成的siRNA是一种新兴的工具，现在广泛应用于这些研究。 不同公司采用多种算法来设计siRNA产品，其在功效，特异性和成本等方面不同。 在此使用来自Dharmacon的siGENOME SMARTpool试剂解释siRNA敲低的示例方案。
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