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Cre-lox-regulated conditional RNA interference from transgenes.
Publication Type:Journal Article
Source:Proc Natl Acad Sci U S A, Volume 101, Issue 28, p.10380-5 (2004)
Keywords:Amino Acid Sequence, Animals, Genetic Vectors, Integrases, Lentivirus, Mice, Mice, Transgenic, Molecular Sequence Data, Nuclear Proteins, Repressor Proteins, RNA Interference, Transgenes, Tumor Suppressor Protein p53, Viral Proteins
<p>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.</p>