CRE recombinase-inducible RNA interference mediated by lentiviral vectors - RNA

CRE recombinase-inducible RNA interference mediated by lentiviral vectors
2004-05-11
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.
Full text is available at http://www.pnas.org/cgi/content/full/101/19/7347