Leber先天性黑朦（LCA）是发病最早、最严重的儿童遗传性视网膜病变，无有效防治措施。RPE65 是其中一个引起LCA的常见致病基因，已发现许多突变位点，但由于缺乏人类疾病靶组织及细胞的研究模型，多数突变位点的致病性与致病机理尚不清楚。围绕RPE65-LCA人类疾病模型缺乏的难题和患者特异RPE65新突变位点c.[200T>G]及c.[430T>G] 是否及如何致病的科学问题，本研究应用先进的hiPSC-3D 类视网膜诱导技术和CRISPR/Cas9基因编辑技术，对上述问题进行系统研究，建立患者特异及RPE65基因型特异的hiPSCs和3D类视网膜疾病模型，并在此模型基础上阐明该杂合突变的致病机制和基因矫正的效果，为LCA患者的精准防治研究提供创新研究平台——类视网膜疾病模型和分子靶点，为基因矫正治疗提供依据。研究有重大科学意义及典范作用，成果水平处于世界领先。

Leber congenital amaurosis (LCA) is the earliest and most serious hereditary retinal degenerative disease, and there is no effective treatment so far. RPE65 is one of the common pathogenic genes causing LCA, and many mutation sites have been identified. However, due to the lack of research models with human diseased tissues and or cells, the pathogenicity and underlying mechanisms of most mutations are still unclear. To solve the above issue and to determine whether and how the two novel compound heterozygous mutations (c. [200T>G] and c. [430T>G]) in RPE65 –LCA patient cause disease, this project aims to 1) establish patient's specific and RPE65 gene phenotype specific disease models in hiPSCs derived retinal organoids using CRISPR/Cas9 technology, 2) clarify the pathogenic mechanisms of these mutations causing LCA, and 3) evaluate the efficacy of the mutation correction. This study would provide molecular targets and an innovative research platform- -retinal organoid models for precision medicine of RPE65-LCA patients, and lay a foundation of clinic gene correction treatment. Therefore, this study is of great significance and plays an exemplary role for other retinal diseases.