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PMID: 15123829 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

CRE recombinase-inducible RNA interference mediated by lentiviral vectors.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 19 ·2004-05-11 ·Pages 7347-51

Tiscornia G, Tergaonkar V, Galimi F, Verma IM

Abstract

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.

MeSH Terms
Base Sequence Cell Line Cloning, Molecular DNA Primers Genetic Vectors Humans Integrases/metabolism Lentivirus/genetics RNA Interference Viral Proteins/metabolism
Chemicals
DNA Primers Viral Proteins Cre recombinase Integrases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Tiscornia Gustavo
Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Tergaonkar Vinay
Galimi Francesco
Verma Inder M
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0402107101
Published
2004-05-11
Epub
2004-00-30
Pages
7347-51
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC409921
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.0402107101 PMC Full Text View journal details
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