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...

Celý popis

Uložené v:

Podrobná bibliografia
Vydané v:	Nature Ročník 428; číslo 6981; s. 431 - 437
Hlavní autori:	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
Médium:	Journal Article
Jazyk:	English
Vydavateľské údaje:	London Nature Publishing Group UK 25.03.2004 Nature Publishing Nature Publishing Group
Predmet:	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
On-line prístup:	Získať plný text
Tagy:	Pridať tag Žiadne tagy, Buďte prvý, kto otaguje tento záznam!

Popis
Shrnutí:	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.
Bibliografia:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 ObjectType-Article-2 ObjectType-Feature-1 content type line 23
ISSN:	0028-0836 1476-4687 1476-4687
DOI:	10.1038/nature02371