Nature Genetics
- 33, 401 - 406 (2003)
Published online: 18 February 2003; Corrected online: 14 May 2007 | doi:10.1038/ng1117
There is a Corrigendum (June 2003) associated with this Letter.
There is a Corrigendum (June 2007) associated with this Letter.
A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interferenceDouglas A Rubinson1, 7, Christopher P Dillon1, 2, 7, Adam V Kwiatkowski1, Claudia Sievers1, 2, 3, Lili Yang4, Johnny Kopinja5, Dina L Rooney6, Mingdi Zhang5, Melanie M Ihrig6, Michael T McManus1, 2, Frank B Gertler1, Martin L Scott5 & Luk Van Parijs1, 21
Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA. 2
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. 3
Free University Berlin, Institute of Biochemistry, Berlin, Germany. 4
California Institute of Technology, Pasadena, California, USA. 5
Biogen Inc., Cambridge, Massachusetts, USA. 6
Division of Comparative Medicine, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA. 7
These authors contributed equally to this work.
Correspondence should be addressed to Luk Van Parijs lukvp@mit.edu 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 retroviruses2,
3,
4,
5,
6,
7,
8,
9,
10. But the resistance of important cell types to transduction by these approaches, both in vitro and in vivo
11, 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.
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