Elucidation of circadian clock machinery by development of an in vivo multi-gene silencing technique

我们以前已经产生了敲除小鼠表达的候选基因的shRNA参与昼夜节律调节使用慢病毒载体。在这项研究中，我们进一步开发了一种技术，在体内表达两种不同的shRNA，并成功地产生了双敲低小鼠。然而，这些小鼠表现出显着的个体差异的敲低效率和沉默效果之间的世代。因此，我们接下来检查了双敲除或细胞类型特异性敲除小鼠。我们的数据表明，在不同的基因敲除小鼠品系异常的昼夜行为节律，并建议参与新的候选基因在昼夜节律钟机制。

We have previously generated knockdown mice expressing shRNA of candidate genes involved in circadian rhythm regulation using a lentiviral vector. In this study, we further developed a technique to express two different shRNAs in vivo and successfully generated double knockdown mice. However, these mice exhibited significant individual differences in the knockdown efficiency and silencing effects among generations. Therefore, we next examined double knockout or cell type-specific knockout mice. Our data demonstrated abnormal circadian behavioral rhythms in different knockout mice strains and suggested involvements of novel candidate genes in the circadian clock machinery.