Second-generation shRNA libraries covering the mouse and human genomes.

Jose M Silva; Mamie Z Li; Ken Chang; Wei Ge; Michael C Golding; Richard J Rickles; Despina Siolas; Guang Hu; Patrick J Paddison; Michael R Schlabach; Nihar Sheth; Jeff Bradshaw; Julia Burchard; Amit Kulkarni; Guy Cavet; Ravi Sachidanandam; W Richard McCombie; Michele A Cleary; Stephen J Elledge; Gregory J Hannon

doi:10.1038/ng1650

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PubMed · 16200065

Second-generation shRNA libraries covering the mouse and human genomes.

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Abstract

Loss-of-function phenotypes often hold the key to understanding the connections and biological functions of biochemical pathways. We and others previously constructed libraries of short hairpin RNAs that allow systematic analysis of RNA interference-induced phenotypes in mammalian cells. Here we report the construction and validation of second-generation short hairpin RNA expression libraries designed using an increased knowledge of RNA interference biochemistry. These constructs include silencing triggers designed to mimic a natural microRNA primary transcript, and each target sequence was selected on the basis of thermodynamic criteria for optimal small RNA performance. Biochemical and phenotypic assays indicate that the new libraries are substantially improved over first-generation reagents. We generated large-scale-arrayed, sequence-verified libraries comprising more than 140,000 second-generation short hairpin RNA expression plasmids, covering a substantial fraction of all predicted genes in the human and mouse genomes. These libraries are available to the scientific community.

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Animals Gene Library Gene Silencing Genome, Human Humans Mice MicroRNAs Plasmids RNA Interference RNA, Small Interfering Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

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Jose M Silva, Mamie Z Li, Ken Chang, Wei Ge, Michael C Golding, Richard J Rickles, Despina Siolas, Guang Hu, Patrick J Paddison, Michael R Schlabach, Nihar Sheth, Jeff Bradshaw, Julia Burchard, Amit Kulkarni, Guy Cavet, Ravi Sachidanandam, W Richard McCombie, Michele A Cleary, Stephen J Elledge, Gregory J Hannon. 2005-10-02. Second-generation shRNA libraries covering the mouse and human genomes.. https://doi.org/10.1038/ng1650

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