A large-scale RNAi screen in human cells identifies new components of the p53 pathway

RNA interference (RNAi) is a powerful new tool with which to perform loss-of-function genetic screens in lower organisms and can greatly facilitate the identification of components of cellular signalling pathways 1 , 2 , 3 . In mammalian cells, such screens have been hampered by a lack of suitable t...

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Bibliographic Details
Published in:Nature Vol. 428; no. 6981; pp. 431 - 437
Main Authors: Berns, Katrien, Hijmans, E. Marielle, Mullenders, Jasper, Brummelkamp, Thijn R., Velds, Arno, Heimerikx, Mike, Kerkhoven, Ron M., Madiredjo, Mandy, Nijkamp, Wouter, Weigelt, Britta, Agami, Reuven, Ge, Wei, Cavet, Guy, Linsley, Peter S., Beijersbergen, Roderick L., Bernards, René
Format: Journal Article
Language:English
Published: London Nature Publishing Group UK 25.03.2004
Nature Publishing
Nature Publishing Group
Subjects:
Biological and medical sciences
Cell cycle
Cell Division
Cell Line, Tumor
Cloning, Molecular
Cyclin-Dependent Kinase Inhibitor p16
Cyclin-Dependent Kinase Inhibitor p21
Cyclins - genetics
Cyclins - metabolism
Down-Regulation
Fibroblasts
Fundamental and applied biological sciences. Psychology
Gene expression
Gene Library
Genes. Genome
Genetic Vectors - genetics
Genetics
Humanities and Social Sciences
Humans
letter
Mammals
Medical screening
Molecular and cellular biology
Molecular genetics
multidisciplinary
Reproducibility of Results
Retroviridae - genetics
Ribonucleic acid
RNA
RNA Interference
RNA, Small Interfering - genetics
RNA, Small Interfering - metabolism
Science
Science (multidisciplinary)
shRNA
Tumor Suppressor Protein p14ARF - metabolism
Tumor Suppressor Protein p53 - genetics
Tumor Suppressor Protein p53 - metabolism
ISSN:0028-0836, 1476-4687, 1476-4687
Online Access:Get full text
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Summary:RNA interference (RNAi) is a powerful new tool with which to perform loss-of-function genetic screens in lower organisms and can greatly facilitate the identification of components of cellular signalling pathways 1 , 2 , 3 . In mammalian cells, such screens have been hampered by a lack of suitable tools that can be used on a large scale. We and others have recently developed expression vectors to direct the synthesis of short hairpin RNAs (shRNAs) that act as short interfering RNA (siRNA)-like molecules to stably suppress gene expression 4 , 5 . Here we report the construction of a set of retroviral vectors encoding 23,742 distinct shRNAs, which target 7,914 different human genes for suppression. We use this RNAi library in human cells to identify one known and five new modulators of p53 -dependent proliferation arrest. Suppression of these genes confers resistance to both p53 -dependent and p19 ARF -dependent proliferation arrest, and abolishes a DNA-damage-induced G1 cell-cycle arrest. Furthermore, we describe siRNA bar-code screens to rapidly identify individual siRNA vectors associated with a specific phenotype. These new tools will greatly facilitate large-scale loss-of-function genetic screens in mammalian cells.
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ISSN:0028-0836
1476-4687
1476-4687
DOI:10.1038/nature02371