基于高通量测序和连锁分析鉴定先天性白内障致病基因

项目名称： 基于高通量测序和连锁分析鉴定先天性白内障致病基因

项目编号： No.81200673

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： 王开杰

作者单位： 首都医科大学

项目金额： 23万元

中文摘要： 先天性白内障是儿童首位致盲性眼病，已有研究表明50%与遗传有关，截止目前已发现18个致病基因与常染色体显性遗传性白内障有关。但是，我们在前期工作中发现目前已知的致病基因仅能解释20%先天性白内障的发生，说明存在更重要的基因在先天性白内障发病中起作用。我们基于2个已排除已知致病基因的先天性白内障家系，通过NimbleGen 液相捕获平台进行外显子组测序，目标外显子区域测序深度50X以上，覆盖度高达99%。本研究拟进一步对测序数据进行高级生物信息学分析筛选候选基因，利用高通量测序和连锁分析相结合的策略，在家系内、另外50个先天性白内障家系及300正常对照对候选基因进行验证。构建真核表达体系，运用细胞生物学手段研究突变对蛋白质溶解性、亚细胞定位及细胞凋亡的影响，对其进行功能验证，最终确定致病基因。研究成果将为阐明先天性白内障发病机制、开展其产前诊断、预防和基因治疗奠定基础。

中文关键词： 先天性白内障；外显子测序；致病基因；；

英文摘要： Congenital cataract is a leading cause of visual disability in children, and approximately half of them are genetically determined. A total of 18 causative genes associated with autosomal dominant congenital cataracts have been identified to date. However, we have previously found that all of the 18 causative genes can only account for 20% of the congenital cataract cases, which indicates that more important genes remain to be discovered. In this study, we captured exomes of eight individuals from two independent Chinese families with congenital cataracts using the platform of Nimblegen exome capture, and performed high-throughput sequencing with a mean coverage of 50×nd sufficient depth to call variants at 99% of each targeted exome. We will exclude the candidate genes by bioinformatics analysis in the next step. The possible causative genes will be identified using a combined strategy of exome sequencing and linkage analysis in the two families. To further confirm it, 300 control individuals of matched geographical ancestry and 50 additional unrelated autosomal dominant congenital cataract families where the known causative gene mutations have been excluded will be examined using Sanger sequencing or MassARRAY. Recombinant wildtype and mutant constructs of the novel causative gene will be synthesized. Triton

英文关键词： congenital cataract；exome sequencing；causitive gene；；