Conditional suppression of cellular genes: lentivirus vector-mediated drug-inducible RNA interference.
RNA interference has emerged as a powerful technique to downregulate the expression of specific genes in cells and in animals, thus opening new perspectives in fields ranging from developmental genetics to molecular therapeutics. Here, we describe a method that significantly expands the potential of RNA interference by permitting the conditional suppression of genes in mammalian cells. Within a lentivirus vector background, we subjected the polymerase III promoter-dependent production of small interfering RNAs to doxycycline-controllable transcriptional repression. The resulting system can achieve the highly efficient and completely drug-inducible knockdown of cellular genes. As lentivirus vectors can stably transduce a wide variety of targets both in vitro and in vivo and can be used to generate transgenic animals, the present system should have broad applications.
Wiznerowicz
Maciej
M
Department of Genetics and Microbiology, CMU, Faculty of Medicine, University of Geneva, 1211 Geneva 4, Switzerland.
Trono
Didier
D
D002460:Cell Line; D004318:Doxycycline / Q000494:pharmacology; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D034622:RNA Interference; D012320:RNA Polymerase III / Q000378:metabolism
D012320:RNA Polymerase III; D004318:Doxycycline