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

Paddison, P. J., Silva, J. M., Conklin, D. S., Schlabach, M., Li, M., Aruleba, S., Balija, V., O'Shaughnessy, A., Gnoj, L., Scobie, K., Chang, K., Westbrook, T., Cleary, M., Sachidanandam, R., McCombie, W. R., Elledge, S. J., Hannon, G. J. (2004) A resource for large-scale RNA-interference-based screens in mammals. Nature, 428 (6981). pp. 427-31. ISSN 1476-4687 (Electronic)

URL: http://www.ncbi.nlm.nih.gov/pubmed/15042091
DOI: 10.1038/nature02370

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.

Item Type: Paper
Uncontrolled Keywords: Animals Cloning Molecular Cysteine Endopeptidases genetics/metabolism Gene Library Genes genetics Genetic Engineering methods Genetic Vectors Humans Mice Multienzyme Complexes genetics metabolism Oligonucleotide Array Sequence Analysis Proteasome Endopeptidase Complex RNA Interference RNA Small Interfering genetics metabolism Reproducibility of Results Substrate Specificity
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNAi
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene silencing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > shRNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > siRNA
CSHL Authors:
Communities: CSHL labs > Hannon lab
CSHL labs > McCombie lab
Watson School > Publications
Depositing User: CSHL Librarian
Date Deposited: 26 Jan 2012 17:19
Last Modified: 19 Sep 2014 15:09
Related URLs:
URI: http://repository.cshl.edu/id/eprint/22456

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