Seibler Jost, Kleinridders Andre, Küter-Luks Birgit, Niehaves Sandra, Brüning Jens C, Schwenk Frieder
Artemis Pharmaceuticals GmbH, Neurather Ring 1, Cologne, Germany.
Nucleic Acids Res. 2007;35(7):e54. doi: 10.1093/nar/gkm122. Epub 2007 Mar 21.
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.
通过表达短发夹（sh）RNA进行RNA干扰为小鼠遗传学中的基因功能分析提供了一种有效方法。然而，目前仍缺乏能够在体内控制基因沉默时间和程度的技术。在此，我们提供了一种普遍适用的系统，用于在小鼠体内对普遍存在的shRNA表达进行时间控制。根据诱导剂强力霉素的剂量，敲低效率可达90%。为了证明我们工具的可行性，通过表达胰岛素受体（Insr）特异性shRNA构建了一个可逆性胰岛素抵抗小鼠模型。诱导后，小鼠在七天内出现严重高血糖。疾病的发作和进展与强力霉素的浓度相关，在撤去诱导剂后不久，表型就会恢复到基线水平。从广泛意义上讲，这种方法将有助于对基因功能和分子疾病机制有新的认识。
