Cre-Lox-Regulated Conditional RNA Interference from Transgenes

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 ha...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2004-07, Vol.101 (28), p.10380-10385
Main Authors: Ventura, Andrea, Meissner, Alexander, Dillon, Christopher P., McManus, Michael, Sharp, Phillip A., Van Parijs, Luk, Jaenisch, Rudolf, Jacks, Tyler
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Biological Sciences
Cell lines
DNA
Embryos
Gene expression
Genetic vectors
Genetic Vectors - genetics
Genetics
Infections
Integrases - genetics
Lentivirus
Lentivirus - genetics
Mathematical vectors
Methylation
Mice
Mice, Transgenic
Molecular Sequence Data
Nuclear Proteins - genetics
Repressor Proteins - genetics
Ribonucleic acid
RNA
RNA Interference
TATA box
Transgenes - genetics
Transgenic animals
Tumor Suppressor Protein p53 - genetics
Viral Proteins - genetics
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Summary: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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0403954101