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

Patrick J. Paddison, Jose M. Silva, Douglas S. Conklin, Mike Schlabach, Mamie Li, Shola Aruleba, Vivekanand Balija, Andy O'Shaughnessy, Lidia Gnoj, Kim Scobie, Kenneth Chang, Thomas Westbrook, Michele Cleary, Ravi Sachidanandam, W. Richard McCombie, Stephen J. Elledge and Gregory J. Hannon ()
Additional contact information
Patrick J. Paddison: Watson School of Biological Sciences
Jose M. Silva: Watson School of Biological Sciences
Douglas S. Conklin: Watson School of Biological Sciences
Mike Schlabach: Howard Hughes Medical Institute, Baylor College of Medicine
Mamie Li: Howard Hughes Medical Institute, Baylor College of Medicine
Shola Aruleba: Watson School of Biological Sciences
Vivekanand Balija: Watson School of Biological Sciences
Andy O'Shaughnessy: Watson School of Biological Sciences
Lidia Gnoj: Watson School of Biological Sciences
Kim Scobie: Watson School of Biological Sciences
Kenneth Chang: Watson School of Biological Sciences
Thomas Westbrook: Howard Hughes Medical Institute, Baylor College of Medicine
Michele Cleary: Rosetta Inpharmatics
Ravi Sachidanandam: Watson School of Biological Sciences
W. Richard McCombie: Watson School of Biological Sciences
Stephen J. Elledge: Howard Hughes Medical Institute, Baylor College of Medicine
Gregory J. Hannon: Watson School of Biological Sciences

Nature, 2004, vol. 428, issue 6981, 427-431

Abstract: Abstract Gene silencing by RNA interference (RNAi) in mammalian cells using small interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) has become a valuable genetic tool1,2,3,4,5,6,7,8,9,10. 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.

Date: 2004
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DOI: 10.1038/nature02370

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