title: A resource for large-scale RNA-interference-based screens in mammals.
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.
authors.affiliation: Cold Spring Harbor Laboratory, Watson School of Biological Sciences, 1 Bungtown Road, Cold Spring Harbor, New York 11724, USA.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Paddison
authors.forename: Jose M
authors.initials: JM
authors.lastname: Silva
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Conklin
authors.forename: Mike
authors.initials: M
authors.lastname: Schlabach
authors.forename: Mamie
authors.initials: M
authors.lastname: Li
authors.forename: Shola
authors.initials: S
authors.lastname: Aruleba
authors.forename: Vivekanand
authors.initials: V
authors.lastname: Balija
authors.forename: Andy
authors.initials: A
authors.lastname: O'Shaughnessy
authors.forename: Lidia
authors.initials: L
authors.lastname: Gnoj
authors.forename: Kim
authors.initials: K
authors.lastname: Scobie
authors.forename: Kenneth
authors.initials: K
authors.lastname: Chang
authors.forename: Thomas
authors.initials: T
authors.lastname: Westbrook
authors.forename: Michele
authors.initials: M
authors.lastname: Cleary
authors.forename: Ravi
authors.initials: R
authors.lastname: Sachidanandam
authors.forename: W Richard
authors.initials: WR
authors.lastname: McCombie
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Elledge
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Hannon
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D005796:Genes / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D046988:Proteasome Endopeptidase Complex; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D015203:Reproducibility of Results; D013379:Substrate Specificity
chemical_list: D009097:Multienzyme Complexes; D034741:RNA, Small Interfering; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex