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Reversible gene knockdown in mice using a tight, inducible shRNA expression system
Authors:Seibler Jost  Kleinridders Andre  Küter-Luks Birgit  Niehaves Sandra  Brüning Jens C  Schwenk Frieder
Affiliation:Jost Seibler, Andre Kleinridders, Birgit Küter-Luks, Sandra Niehaves, Jens C. Brüning, and Frieder Schwenk
Abstract:RNA interference through expression of short hairpin (sh)RNAs provides an efficient approach for gene function analysis in mouse genetics. Techniques allowing to control time and degree of gene silencing in vivo, however, are still lacking. Here we provide a generally applicable system for the temporal control of ubiquitous shRNA expression in mice. Depending on the dose of the inductor doxycycline, the knockdown efficiency reaches up to 90%. To demonstrate the feasibility of our tool, a mouse model of reversible insulin resistance was generated by expression of an insulin receptor (Insr)-specific shRNA. Upon induction, mice develop severe hyperglycemia within seven days. The onset and progression of the disease correlates with the concentration of doxycycline, and the phenotype returns to baseline shortly after withdrawal of the inductor. On a broad basis, this approach will enable new insights into gene function and molecular disease mechanisms.
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