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Advanced Review RNAi screening new approaches understandings and organisms Stephanie E Mohr1 and Norbert Perrimon2 RNA interference RNAi leads to sequence specific knockdown of gene function The approach can be used in large scale screens to interrogate function in various model organisms and an increasing number of other species Genome scale RNAi screens are routinely performed in cultured or primary cells or in vivo in organisms such as C elegans High throughput RNAi screening is benefitting from the development of sophisticated new instrumentation and software tools for collecting and analyzing data including high content image data The results of large scale RNAi screens have already proved useful leading to new understandings of gene function relevant to topics such as infection cancer obesity and aging Nevertheless important caveats apply and should be taken into consideration when developing or interpreting RNAi screens Some level of false discovery is inherent to high throughput approaches and specific to RNAi screens false discovery due to off target effects OTEs of RNAi reagents remains a problem The need to improve our ability to use RNAi to elucidate gene function at large scale and in additional systems continues to be addressed through improved RNAi library design development of innovative computational and analysis tools and other approaches 2011 John Wiley Sons Ltd WIREs RNA 2011 DOI 10 1002 wrna 110 INTRODUCTION R NA interference RNAi is a conserved endogenous activity1 that can be harnessed as a tool for functional genomics studies 2 8 With RNAi genespecific reagents are introduced into cells triggering knockdown or reduction of gene function via sequence specific degradation and translational interference of mRNA transcripts RNAi screening provides a powerful reverse genetic approach to largescale functional analysis in cultured cells and in an increasing number of in vivo systems Like genetic screening RNAi screening allows for identification of

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