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.
(March 2004)
A resource for large-scale RNA-interference-based screens in mammals.
Nature, 428 (6981).
pp. 427-31.
ISSN 1476-4687 (Electronic)
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: |
Aruleba, Shola
Balija, Vivekanand
Chang, Kenneth
Conklin, Douglas S.
Gnoj, Lidia
Hannon, Gregory J.
McCombie, W. Richard
O’Shaughnessy, Andrew L.
Paddison, Patrick J.
Sachidanandam, Ravi
Scoble, Kim
Silva, Jose M.
|
| Communities: |
CSHL labs > Hannon lab
CSHL labs > McCombie lab
School of Biological Sciences > Publications |
| Depositing User: |
CSHL Librarian
|
| Date: |
25 March 2004 |
| Date Deposited: |
26 Jan 2012 17:19 |
| Last Modified: |
19 Sep 2014 15:09 |
| Related URLs: |
|
| URI: |
https://repository.cshl.edu/id/eprint/22456 |
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