A resource for large-scale RNA-interference-based screens in mammals
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 libr...
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| Published in: | Nature Vol. 428; no. 6981; pp. 427 - 431 |
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| Main Authors: | , , , , , , , , , , , , , , , , |
| Format: | Journal Article |
| Language: | English |
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London
Nature Publishing
25-03-2004
Nature Publishing Group |
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| Online Access: | Get full text |
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| 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. |
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| AbstractList | 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. 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. [PUBLICATION ABSTRACT] |
| Audience | Academic |
| Author | Li, Mamie Elledge, Stephen J Sachidanandam, Ravi O'Shaughnessy, Andy Chang, Kenneth Hannon, Gregory J Balija, Vivekanand Scobie, Kim Westbrook, Thomas Silva, Jose M Schlabach, Mike Gnoj, Lidia Cleary, Michele Paddison, Patrick J Conklin, Douglas S Aruleba, Shola Richard McCombie, W |
| Author_xml | – sequence: 1 givenname: Gregory J surname: Hannon fullname: Hannon, Gregory J – sequence: 2 givenname: Patrick J surname: Paddison fullname: Paddison, Patrick J – sequence: 3 givenname: Jose M surname: Silva fullname: Silva, Jose M – sequence: 4 givenname: Douglas S surname: Conklin fullname: Conklin, Douglas S – sequence: 5 givenname: Mike surname: Schlabach fullname: Schlabach, Mike – sequence: 6 givenname: Mamie surname: Li fullname: Li, Mamie – sequence: 7 givenname: Shola surname: Aruleba fullname: Aruleba, Shola – sequence: 8 givenname: Vivekanand surname: Balija fullname: Balija, Vivekanand – sequence: 9 givenname: Andy surname: O'Shaughnessy fullname: O'Shaughnessy, Andy – sequence: 10 givenname: Lidia surname: Gnoj fullname: Gnoj, Lidia – sequence: 11 givenname: Kim surname: Scobie fullname: Scobie, Kim – sequence: 12 givenname: Kenneth surname: Chang fullname: Chang, Kenneth – sequence: 13 givenname: Thomas surname: Westbrook fullname: Westbrook, Thomas – sequence: 14 givenname: Michele surname: Cleary fullname: Cleary, Michele – sequence: 15 givenname: Ravi surname: Sachidanandam fullname: Sachidanandam, Ravi – sequence: 16 givenname: W surname: Richard McCombie fullname: Richard McCombie, W – sequence: 17 givenname: Stephen J surname: Elledge fullname: Elledge, Stephen J |
| BackLink | http://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15637277$$DView record in Pascal Francis https://www.ncbi.nlm.nih.gov/pubmed/15042091$$D View this record in MEDLINE/PubMed |
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| CODEN | NATUAS |
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| Copyright | 2004 INIST-CNRS COPYRIGHT 2004 Nature Publishing Group Copyright Macmillan Journals Ltd. Mar 25, 2004 |
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| References | Hannon, G. J. (b1) 2002; 418 Paul, C. P., Good, P. D., Winer, I., Engelke, D. R. (b9) 2002; 20 Lum, L. (b2) 2003; 299 Bochtler, M., Ditzel, L., Groll, M., Hartmann, C., Huber, R. (b17) 1999; 28 Fraser, A. G. (b5) 2000; 408 Ghoda, L., Sidney, D., Macrae, M., Coffino, P. (b14) 1992; 12 Kim, S. Y., Herbst, A., Tworkowski, K. A., Salghetti, S. E., Tansey, W. P. (b19) 2003; 11 Coux, O. (b18) 2003; 31 Paddison, P. J., Caudy, A. A., Bernstein, E., Hannon, G. J., Conklin, D. S. (b6) 2002; 16 Heinemeyer, W., Fischer, M., Krimmer, T., Stachon, U., Wolf, D. H. (b16) 1997; 272 Birrell, G. W., Giaever, G., Chu, A. M., Davis, R. W., Brown, J. M. (b11) 2001; 98 Lee, S. S. (b3) 2003; 33 Winzeler, E. A. (b13) 1999; 285 Hemann, M. T. (b7) 2003; 33 Lois, C., Hong, E. J., Pease, S., Brown, E. J., Baltimore, D. (b10) 2002; 295 Chen, P., Hochstrasser, M. (b15) 1996; 86 Paddison, P. J., Hannon, G. J. (b8) 2003; 5 Gonczy, P. (b4) 2000; 408 Giaever, G. (b12) 2002; 418 15042071 - Nature. 2004 Mar 25;428(6981):375-8 |
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| Title | A resource for large-scale RNA-interference-based screens in mammals |
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