Abstract
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.
Keywords: Pooled RNAi screen, molecular tag, barcode, shRNA library, microarray, half hairpin
Current Genomics
Title: Pooled RNAi Screens - Technical and Biological Aspects
Volume: 11 Issue: 3
Author(s): M. Boettcher and J.D. Hoheisel
Affiliation:
Keywords: Pooled RNAi screen, molecular tag, barcode, shRNA library, microarray, half hairpin
Abstract: 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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Cite this article as:
Boettcher M. and Hoheisel J.D., Pooled RNAi Screens - Technical and Biological Aspects, Current Genomics 2010; 11 (3) . https://dx.doi.org/10.2174/138920210791110988
DOI https://dx.doi.org/10.2174/138920210791110988 |
Print ISSN 1389-2029 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5488 |
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