CRE recombinase-inducible RNA interference mediated by lentiviral vectors.
Recently, several systems designed to trigger RNA interference by using small hairpin RNA driven by polymerase III promoters have been described. Here, we report a lentiviral-mediated small interfering RNA delivery system that can be induced by CRE recombinase. The system consists of a lentiviral vector carrying a mouse U6 promoter that is separated from a small hairpin RNA by a random DNA stuffer sequence flanked by modified loxP sites. The silencing cassette is not expressed until activated by addition of CRE recombinase delivered by a lentiviral vector. We have used this system to show specific down-regulation of GFP and two endogenous genes (the tumor suppressor p53 and the NF-kappaB transcription factor subunit p65) in vitro. Furthermore, down-regulation of both p53 and p65 resulted in the expected effect on downstream genes and cellular phenotype. We foresee multiple applications of this system both in vitro and in vivo to down-regulate specific targets in a tissue-specific and localized manner.
Tiscornia
Gustavo
G
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Tergaonkar
Vinay
V
Galimi
Francesco
F
Verma
Inder M
IM
D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D005822:Genetic Vectors; D006801:Humans; D019426:Integrases / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics*; D034622:RNA Interference; D014764:Viral Proteins / Q000378:metabolism*
D017931:DNA Primers; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases