视网膜色素变性（RP）是由基因变异引起的致盲性眼病。其分子致病机理极为复杂，近半数患者病因仍不明了。申请人前期通过全外显子测序在RP家系中初步发现了新型致病基因CEP250，本课题将针对CEP250的分子致病机理和基因干预实验展开研究。首先进行大样本量筛查，拟在已收集的1044例RP中筛查CEP250以研究该基因的突变频谱；基于细胞实验探究已发现的无义突变对细胞定位和蛋白功能的影响；建立斑马鱼模型，利用斑马鱼视动反应和行为学分析系统检测评估CEP250基因缺陷对斑马鱼视觉系统发育和视觉功能的影响；构建与先证者相同无义突变的敲入小鼠模型，探究CEP250基因缺陷对于纤毛发育的影响，以及对小鼠视网膜的结构形态和功能的影响；再进一步利用基因及蛋白表达和基因修复等手段探究其分子致病机理及基因修复的治疗效果，从而为RP的精确诊断和治疗提供关键的科学依据。

Retinitis pigmentosa (RP), caused by genetic defects, can lead to eventual blindness. Despite the fact that more than 70 genes are known to cause RP, approximately 40% cases remain to be solved due to the enormous genetic heterogeneity. Based on my previous researches on the genetic basis of RP, I identified CEP250 gene as a novel RP gene through whole exome sequencing. In this study, we will screen the CEP250 in a large RP cohort, including 1044 cases, to evaluate the contribution to RP. The effect of mutant would be tested according to subcellular and co-immunoprecipitation. Subsequently, we will develop a cep250-knockdown zebrafish model to test the function of cep250 in zebrafish. Furthermore, we will develop a Cep250-knockin mice model to determine the molecular mechanism of cilia and retina, and the efficiency of gene replacement. Elucidating disease mechanism and testing the gene replacement of CEP250 could benefit the genetic diagnosis and gene therapy of RP patients.