title: A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference.
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.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Rubinson
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Dillon
authors.forename: Adam V
authors.initials: AV
authors.lastname: Kwiatkowski
authors.forename: Claudia
authors.initials: C
authors.lastname: Sievers
authors.forename: Lili
authors.initials: L
authors.lastname: Yang
authors.forename: Johnny
authors.initials: J
authors.lastname: Kopinja
authors.forename: Dina L
authors.initials: DL
authors.lastname: Rooney
authors.forename: Mingdi
authors.initials: M
authors.lastname: Zhang
authors.forename: Melanie M
authors.initials: MM
authors.lastname: Ihrig
authors.forename: Michael T
authors.initials: MT
authors.lastname: McManus
authors.forename: Frank B
authors.initials: FB
authors.lastname: Gertler
authors.forename: Martin L
authors.initials: ML
authors.lastname: Scott
authors.forename: Luk
authors.initials: L
authors.lastname: Van Parijs
mesh_terms: D000818:Animals; D001483:Base Sequence; D016827:CD8 Antigens / Q000235:genetics; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry / Q000235:genetics; D034622:RNA Interference; D015375:Receptors, Interleukin-2 / Q000235:genetics; D013234:Stem Cells
chemical_list: D016827:CD8 Antigens; D008164:Luminescent Proteins; D015375:Receptors, Interleukin-2; D049452:Green Fluorescent Proteins; D012313:RNA