Pooled RNAi screens-technical and biological aspects

M Boettcher, JD Hoheisel - Current genomics, 2010 - ingentaconnect.com
M Boettcher, JD Hoheisel
Current genomics, 2010ingentaconnect.com
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 …
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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