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RNAi screening: new approaches, understandings, and organisms

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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. WIREs RNA 2012, 3:145–158. doi: 10.1002/wrna.110

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RNA Methods
RNA Methods > RNA Analyses in Cells
Regulatory RNAs/RNAi/Riboswitches > RNAi: Mechanisms of Action

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