基因敲除技术研究ezrin维持晶状体结构的机制

项目名称： 基因敲除技术研究ezrin维持晶状体结构的机制

项目编号： No.81200667

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学二处

项目作者： 周南

作者单位： 哈尔滨医科大学

项目金额： 23万元

中文摘要： 本课题申请人通过外显子组测序发现了基因EZR的突变可能导致一个四代中国人先天性核性白内障的发生，这是世界上对该基因是先天性白内障可疑致病基因的首次报道，但目前仍需通过EZR基因敲除鼠进一步验证该基因与先天性白内障的关系。EZR基因所编译膜细胞骨架连接蛋白ezrin是连接细胞膜和细胞骨架之间ERM蛋白家族成员之一。它与跨膜蛋白和细胞骨架蛋白的相互连接对膜结构域的形成有着极其重要的作用，因此ezrin作为连接蛋白可能对维持晶状体纤维细胞有序结构，保持晶状体的透明性有着十分重要的意义，但目前对其在维持晶状体透明性的具体机制尚不清楚。本课题拟利用条件性基因敲除技术，建立晶状体EZR基因敲除小鼠模型，从而利用光学显微镜、扫描电镜、冷冻蚀刻电镜、免疫组化及分子生物学方式观察基因敲除后晶状体纤维的有序组织结构的变化和晶状体透明性的改变，从而证实该基因是先天性白内障的致病基因。

中文关键词： EZR基因；基因突变；斑马鱼；先天性白内障；

英文摘要： As the next-generation sequencing technology, exome sequencing has been valued and applied widely in the field of genetics, including the genetic eye disorders. It has been successfully used in our last research. A mutant gene EZR was found in a four generation Chinese congenital cataract family. This is the first time that the EZR gene has been reported as a suspicious congenital cataract pathogenic cause in the world. However, this result still needs knockout technology to confirm further. Therefore, in this study, we will perform light microscopy, scanning electron microscopy, freeze-fracture transmission electron microscopy immunohistochemistry and molecular biological technology on the EZR knockout mouse to understand the protein ezrin well in the lens. The protein ezrin, one of the ERM proteins (ezrin, radixin and moesin) encoded by gene EZR,mainly presents in the young elongating cortical fiber cells and localizes to the plasma membranes of the lens. In recent years, the studies focused on the distribution and functions of ezrin in the lens reveals that ERM protein family especially ezrin plays a crucial role in the development of the lens, and its reduction or disfuction would potentially cause the disorder of the fiber cells which inducing the opacification of the lens. However, the exactly mechanism o

英文关键词： EZR；gene mutation；zebrafish；congenital cataract；