A resource for large-scale RNA-interference-based screens in mammals

Gene silencing by RNA interference (RNAi) in mammalian cells using small interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) has become a valuable genetic tool. Here, we report the construction and application of a shRNA expression library targeting 9,610 human and 5,563 mouse genes. This libr...

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Bibliographic Details
Published in:	Nature Vol. 428; no. 6981; pp. 427 - 431
Main Authors:	Hannon, Gregory J, Paddison, Patrick J, Silva, Jose M, Conklin, Douglas S, Schlabach, Mike, Li, Mamie, Aruleba, Shola, Balija, Vivekanand, O'Shaughnessy, Andy, Gnoj, Lidia, Scobie, Kim, Chang, Kenneth, Westbrook, Thomas, Cleary, Michele, Sachidanandam, Ravi, Richard McCombie, W, Elledge, Stephen J
Format:	Journal Article
Language:	English
Published:	London Nature Publishing 25-03-2004 Nature Publishing Group
Subjects:	Animals Biological and medical sciences Cloning, Molecular Cysteine Endopeptidases - genetics Cysteine Endopeptidases - metabolism Fundamental and applied biological sciences. Psychology Gene expression Gene Library Genes - genetics Genes. Genome Genetic Engineering - methods Genetic research Genetic resources Genetic Vectors Genetics Humans Mammals Mice Molecular and cellular biology Molecular genetics Multienzyme Complexes - genetics Multienzyme Complexes - metabolism Oligonucleotide Array Sequence Analysis Proteasome Endopeptidase Complex Reproducibility of Results Ribonucleic acid RNA RNA Interference RNA, Small Interfering - genetics RNA, Small Interfering - metabolism Rodents shRNA Substrate Specificity
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Summary:	Gene silencing by RNA interference (RNAi) in mammalian cells using small interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) has become a valuable genetic tool. Here, we report the construction and application of a shRNA expression library targeting 9,610 human and 5,563 mouse genes. This library is presently composed of about 28,000 sequence-verified shRNA expression cassettes contained within multi-functional vectors, which permit shRNA cassettes to be packaged in retroviruses, tracked in mixed cell populations by means of DNA 'bar codes', and shuttled to customized vectors by bacterial mating. In order to validate the library, we used a genetic screen designed to report defects in human proteasome function. Our results suggest that our large-scale RNAi library can be used in specific, genetic applications in mammals, and will become a valuable resource for gene analysis and discovery.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0028-0836 1476-4687
DOI:	10.1038/nature02370