Knock-down mouse, a novel gene engineering mouse for use as a disease model

在本研究中，我们试图构建一种新型的基因工程小鼠，该小鼠包含四环素调控基因表达系统（Tet-OFF和Tet-ON）和由该开关系统驱动的利比赛酶基因。首先，我们构建了一个抗胰岛素核酶，并使用合成的胰岛素RNA和核酶进行体外切割实验。该核酶对RNA具有体外裂解活性。该核酶在M6胰岛素瘤细胞转染系统中也显示出抗胰岛素活性。制备了用于血管内皮生长因子（VEGF）和活化K-ras基因RNA的核酶，并在细胞转化和体外裂解实验系统中显示了裂解活性。此外，转染抗vegf或抗k -ras核酶的肿瘤细胞在裸鼠中的致瘤活性降低。利用Tet-OFF和Tet-ON系统分别建立了转染rusiferase基因的转基因小鼠。老鼠很兴奋。

In this research, we attempt to create a novel gene engineering mouse which contains the target gene expression-switch such as tetracycline regulated gene expression system (Tet-OFF and Tet-ON) and a gene for ribizyme driven by the switch system. First, we constructed an anti-insulin ribozyme, and examined in vitro cleaving assay using synthesized insulin RNA and the ribozyme. The ribozyme showed in vitro cleavage activity to the RNA.The ribozyme also showed anti-insulin activity in M6 insulinoma cell transfectant system. Another ribozymes were created for VEGF (vascular endotherial growth factor) and activated K-ras gene RNA, and they showed cleaving activity in cell transformant and in vitro cleavage assay system.Also, tumor cells transfected by the anti-VEGF or anti-K-ras ribozyme showed reduced tumorigenic activity in nude mouse. Transgenic mice for rusiferase gene driven by the Tet-OFF and Tet-ON system were created. The mice were under exanimation.