Pooled RNAi Screens - Technical and Biological Aspects.

Pooled RNAi Screens - Technical and Biological Aspects.
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DOI:
10.2174/138920210791110988
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发表时间:
2010-05
期刊:
影响因子:
2.6
通讯作者:
Hoheisel JD
Hoheisel JD
中科院分区:
生物学4区
文献类型:
--
作者:
Boettcher M;Hoheisel JD

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RNA干扰(RNAi)筛选最近成为研究哺乳动物细胞基因功能的一种令人兴奋的新工具。为了促进这些研究,覆盖整个人类转录组的短发夹RNA (shRNA)表达文库已经商品化。为了充分利用这种大规模shRNA文库的潜力,已经开发出基于微阵列的方法来分析复杂的聚合RNAi筛选。在微阵列分析方面,不同的研究小组采用了不同的策略,这在很大程度上受到所使用的shRNA文库的影响。在这篇综述中,我们比较了三种主要的shRNA表达文库,重点讨论了它们在基于微阵列的汇总筛选分析中的适用性。我们分析和比较了以前用于执行集合RNAi筛选的方法,并指出了它们的优点和局限性。
RNA interference (RNAi) screens have recently emerged as an exciting new tool for studying gene function in mammalian cells. In order to facilitate those studies, short hairpin RNA (shRNA) expression libraries covering the entire human transcriptome have become commercially available. To make use of the full potential of such large-scale shRNA libraries, microarray-based methods have been developed to analyze complex pooled RNAi screens. In terms of microarray analysis, different strategies have been pursued by different research groups, largely influenced by the employed shRNA library. In this review, we compare the three major shRNA expression libraries with a focus on their suitability for a microarray-based analysis of pooled screens. We analyze and compare approaches previously used to perform pooled RNAi screens and point out their advantages as well as limitations.
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