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Single copy shRNA configuration for ubiquitous gene knockdown in mice

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Item Type:Article
Title:Single copy shRNA configuration for ubiquitous gene knockdown in mice
Creators Name:Seibler, J., Küter-Luks, B., Kern, H., Streu, S., Plum, L., Mauer, J., Kühn, R., Bruening, J.C. and Schwenk, F.
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.
Keywords:beta-Galactosidase, Cell Surface Receptors, Cultured Cells, Firefly Luciferases, Gene Dosage, Genetic Promoter Regions, Insertional Mutagenesis, Leptin Receptors, Proteins, RNA Interference, Recombinases, Small Interfering RNA, Transgenes, Untranslated RNA, Animals, Mice
Source:Nucleic Acids Research
Publisher:Oxford University Press
Page Range:e67
Date:14 April 2005
Official Publication:https://doi.org/10.1093/nar/gni065
PubMed:View item in PubMed

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