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PMID: 15240889 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cre-lox-regulated conditional RNA interference from transgenes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 28 ·2004-07-13 ·Pages 10380-5

Ventura A, Meissner A, Dillon CP, McManus M, Sharp PA, Van Parijs L, Jaenisch R, Jacks T

Abstract

We have generated two lentiviral vectors for conditional, Cre-lox-regulated, RNA interference. One vector allows for conditional activation, whereas the other permits conditional inactivation of short hairpin RNA (shRNA) expression. The former is based on a strategy in which the mouse U6 promoter has been modified by including a hybrid between a LoxP site and a TATA box. The ability to efficiently control shRNA expression by using these vectors was shown in cell-based experiments by knocking down p53, nucleophosmin and DNA methyltransferase 1. We also demonstrate the usefulness of this approach to achieve conditional, tissue-specific RNA interference in Cre-expressing transgenic mice. Combined with the growing array of Cre expression strategies, these vectors allow spatial and temporal control of shRNA expression in vivo and should facilitate functional genetic analysis in mammals.

MeSH Terms
Amino Acid Sequence Animals Genetic Vectors/genetics Integrases/genetics Lentivirus/genetics Mice Mice, Transgenic Molecular Sequence Data Nuclear Proteins/genetics Nucleophosmin RNA Interference Repressor Proteins/genetics Transgenes/genetics Tumor Suppressor Protein p53/genetics Viral Proteins/genetics
Chemicals
DMAP1 protein, human Dmap1 protein, mouse NPM1 protein, human Nuclear Proteins Repressor Proteins Tumor Suppressor Protein p53 Viral Proteins Nucleophosmin Cre recombinase Integrases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Ventura Andrea
Department of Biology, Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Meissner Alexander
Dillon Christopher P
McManus Michael
Sharp Phillip A
Van Parijs Luk
Jaenisch Rudolf
Jacks Tyler
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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.0403954101
Published
2004-07-13
Epub
2004-00-06
Pages
10380-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC478580
Subset
IM
Grants
NIGMS NIH HHS · R37 GM034277 · United States
NIGMS NIH HHS · R37-GM34277 · United States
External Links
PubMed source DOI: 10.1073/pnas.0403954101 PMC Full Text View journal details
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