Production of inv (Inversion of embryonic turning) gene knockout mouse and development of disease related animal models

我们根据inv突变小鼠的信息确定了决定左右不对称和调节肾脏发育的inv基因(自然，395,1998)。小鼠inv基因编码1062个氨基酸的蛋白，包含15个串联重复的Ankyrin基序。然而，inv基因的功能特性和基因表达调控因子尚不清楚，本课题构建了inv基因的靶向载体，用于培育inv基因敲除小鼠。同时，利用过表达和RNAi技术分析了inv蛋白在基因表达水平上的调控作用。用基因芯片技术分析通过基因芯片技术改变inv基因表达水平的肾小管上皮细胞。与基质结构蛋白相关的几个基因表达下调，修复相关基因表达上调。此外，我们还鉴定和分析了一些与inv密切相关的基因，如CD2AP或转录因子maf。在不久的将来，我们将对我们在本项目中建立的基因敲除小鼠进行分析，试图揭示inv蛋白的基本作用和功能。

We identified the inv gene which decides the left and right asymmetry and regulates kidney development based on the information of the inv mutant mouse (Nature 395, 1998). The mouse inv gene encodes 1062 amino acid protein containing 15 tandem repeats of the ankyrin motif. However, functional properties and the modulator of gene expression of inv have been unclear.In this project, we constructed the targeting vector of inv gene for producing inv knockout mouse. Simultaneously, we analyzed the function of inv protein by modulation of gene expression level using overexpression and RNAi technique. The cultured renal tubular cell in which the expression level of inv mRNA was modified by transfection or siRNA and the alternation of gene expression profile was analyzed by microarray system. Several genes related to structural protein of the matrix were downregulated, and in contrast, repairing related genes were upregulated. Moreover, we identified and analyzed some genes closely related to the inv, such as CD2AP or transcription factor mafs.Near future, we analyzed knockout mouse that we established in this project and will try to reveal the essential role and function of inv protein.