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  • Title: Single copy shRNA configuration for ubiquitous gene knockdown in mice.
    Author: Seibler J, Küter-Luks B, Kern H, Streu S, Plum L, Mauer J, Kühn R, Brüning JC, Schwenk F.
    Journal: Nucleic Acids Res; 2005 Apr 14; 33(7):e67. PubMed ID: 15831785.
    Abstract:
    RNA interference through the expression of small hairpin RNA (shRNA) molecules has become a very promising tool in reverse mouse genetics as it may allow inexpensive and rapid gene function analysis in vivo. However, the prerequisites for ubiquitous and reproducible shRNA expression are not well defined. Here we show that a single copy shRNA-transgene can mediate body-wide gene silencing in mice when inserted in a defined locus of the genome. The most commonly used promoters for shRNA expression, H1 and U6, showed a comparably broad activity in this configuration. Taken together, the results define a novel approach for efficient interference with expression of defined genes in vivo. Moreover, we provide a rapid strategy for the production of gene knockdown mice combining recombinase mediated cassette exchange and tetraploid blastocyst complementation approaches.
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