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

Author

Listed:
  • Patrick J. Paddison

    (Watson School of Biological Sciences)

  • Jose M. Silva

    (Watson School of Biological Sciences)

  • Douglas S. Conklin

    (Watson School of Biological Sciences
    University at Albany)

  • Mike Schlabach

    (Howard Hughes Medical Institute, Baylor College of Medicine
    Harvard Partners Center for Genetics and Genomics, Harvard Medical School)

  • 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
    Harvard Partners Center for Genetics and Genomics, Harvard Medical School)

  • 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
    Harvard Partners Center for Genetics and Genomics, Harvard Medical School)

  • Gregory J. Hannon

    (Watson School of Biological Sciences)

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.

Suggested Citation

  • 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 Westbro, 2004. "A resource for large-scale RNA-interference-based screens in mammals," Nature, Nature, vol. 428(6981), pages 427-431, March.
    • Handle: RePEc:nat:nature:v:428:y:2004:i:6981:d:10.1038_nature02370
    DOI: 10.1038/nature02370
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