Inducible and reversible gene silencing by stable integration of an shRNA-encoding lentivirus in transgenic rats
Marco J Herold, Jens van den Brandt, Jost Seibler, Holger M Reichardt
Proceedings of the National Academy of Sciences of the United States of America | NATL ACAD SCIENCES | Published : 2008
Currently, tools to generate loss-of-function mutations in rats are limited. Therefore, we have developed a lentiviral single-vector system for the temporal control of ubiquitous shRNA expression. Here, we report transgenic rats carrying an insulin receptor-specific shRNA transcribed from a regulatable promoter and identified by concomitant EGFP expression. In the absence of the inducer doxycycline (Dox), we observed no siRNA expression. However, Dox treatment at very low concentrations led to a rapid induction of the siRNA and ablation of INSR protein expression. As anticipated, blood glucose levels increased, whereas insulin signaling and glucose regulation were impaired. Importantly, this..View full abstract
We thank Christian Bauer and Amina Bassibas for expert technical help, Alex Delbridge for critical reading of the manuscript, and Thomas Hunig for scientific discussion and advice. This work was supported by a grant from the German Ministry for Science and Education and funds by TaconicArtemis GmbH. J.S. is an employee of TaconicArtemis GmbH and ensures that declared interests did not influence the objectivity, integrity, or perceived value of this article.