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A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference

Rubinson, Douglas A. and Dillon, Christopher P. and Kwiatkowski, Adam V. and Sievers, Claudia and Yang, Lili and Kopinja, Johnny and Rooney, Dina L. and Zhang, Mingdi and Ihrig, Melanie M. and McManus, Michael T. and Gertler, Frank B. and Scott, Martin L. and Van Parijs, Luk (2003) A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nature Genetics, 33 (3). pp. 401-406. ISSN 1061-4036. http://resolver.caltech.edu/CaltechAUTHORS:20150408-081455469

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Abstract

RNA interference (RNAi) has recently emerged as a specific and efficient method to silence gene expression in mammalian cells either by transfection of short interfering RNAs (siRNAs; ref. 1) or, more recently, by transcription of short hairpin RNAs (shRNAs) from expression vectors and retroviruses. But the resistance of important cell types to transduction by these approaches, both in vitro and in vivo, has limited the use of RNAi. Here we describe a lentiviral system for delivery of shRNAs into cycling and non-cycling mammalian cells, stem cells, zygotes and their differentiated progeny. We show that lentivirus-delivered shRNAs are capable of specific, highly stable and functional silencing of gene expression in a variety of cell types and also in transgenic mice. Our lentiviral vectors should permit rapid and efficient analysis of gene function in primary human and animal cells and tissues and generation of animals that show reduced expression of specific genes. They may also provide new approaches for gene therapy.


Item Type:Article
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URLURL TypeDescription
http://sc.doi.org/10.1038/ng1117DOIArticle
http://www.nature.com/ng/journal/v33/n3/full/ng1117.htmlPublisherArticle
Additional Information:© 2007 Nature Publishing Group. Published online: 18 February 2003; Corrected online: 14 May 2007. We would like to thank J. Bear, J. Chen, P. Sharp, C. Lois and D. Baltimore for their advice and D. Chojnacky, A. Antov, K. Layer, B. Haines, G. Paradis and the Flow Cytometry facility for their technical support. This study was supported by a grant from the David Koch Cancer Research Fund, a Career Development Award from the Arthritis Foundation and the Juvenile Diabetes Foundation (to L.V.P.), a grant from the US National Institutes of Health and a W M Keck Distinguished Young Scholar Award (to F.B.G.). D.A.R. is supported by the Medical Scientist Training Program. C.P.D. is a Howard Hughes Medical Institute Predoctoral Fellow. M.T.M. is a fellow of the Cancer Research Institute. A.V.K. is supported by an Anna Fuller Predoctoral Scholarship.
Funders:
Funding AgencyGrant Number
David Koch Cancer Research FundUNSPECIFIED
Arthritis FoundationUNSPECIFIED
Juvenile Diabetes FoundationUNSPECIFIED
NIHUNSPECIFIED
W. M. Keck FoundationUNSPECIFIED
UCLA-Caltech Medical Scientist Training ProgramUNSPECIFIED
Anna Fuller Predoctoral ScholarshipUNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Cancer Research InstituteUNSPECIFIED
Record Number:CaltechAUTHORS:20150408-081455469
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20150408-081455469
Official Citation:A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference pp401 - 406 Douglas A Rubinson, Christopher P Dillon, Adam V Kwiatkowski, Claudia Sievers, Lili Yang, Johnny Kopinja, Dina L Rooney, Mingdi Zhang, Melanie M Ihrig, Michael T McManus, Frank B Gertler, Martin L Scott & Luk Van Parijs Published online: 18 February 2003 | doi:10.1038/ng1117
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:56461
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:08 Apr 2015 15:41
Last Modified:08 Apr 2015 15:41

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