Skip to main navigation Skip to search Skip to main content

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

  • Patrick J. Paddison
  • , Jose M. Silva
  • , Douglas S. Conklin
  • , Mlke Schlabach
  • , Mamie Li
  • , Shola Aruleba
  • , Vivekanand Balija
  • , Andy O'Shaughnessy
  • , Lidia Gnoj
  • , Kim Scoble
  • , Kenneth Chang
  • , Thomas Westbrook
  • , Michele Cleary
  • , Ravi Sachidanandam
  • , W. Rlchard McComble
  • , Stephen J. Elledge
  • , Gregory J. Hannon

Research output: Contribution to journalArticlepeer-review

628 Scopus citations

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 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.

Original languageEnglish
Pages (from-to)427-431
Number of pages5
JournalNature
Volume428
Issue number6981
DOIs
StatePublished - 25 Mar 2004
Externally publishedYes

Fingerprint

Dive into the research topics of 'A resource for large-scale RNA-interference-based screens in mammals'. Together they form a unique fingerprint.

Cite this