纳米囊泡介导的CRISPR/Cas9系统联合多组学测序技术对GJB2缺陷导致的常染色体隐性非综合征性耳聋的基因修复的研究

非综合征性耳聋（NSHI）关键基因GJB2突变形式主要是单核苷酸缺失和点突变，由此造成Cx26缝隙连接蛋白的错译和无功能，但除少数GJB2 突变外，突变型和表型之间并无明显的相关性，突变功能改变与表型之间也没有明确的关系。为了解决传统外显子组测序无法满足对其发病机制深入研究的要求，本课题组采用来源于GJB2基因不同突变型的Cx26-NSHI患者的转录组、翻译组及表观组测序数据进行整合分析，将它们之间线性或非线性关系等联系到一起，有效去除单个环节的随机事件，从新视角研究GJB2基因突变型与临床表型之间内在的分子机制。同时为了修复GJB2基因突变导致的Cx26缺陷，本课题组构建GJB2基因缺陷转基因小鼠模型，采用纳米囊泡介导的基因编辑技术针对突变制造纠正来恢复或改善其功能。通过对GJB2基因突变导致的Cx26-NSHI上游的基因检测到疾病建模再到下游基因治疗，形成整体研究思路。

The main forms of GJB2 mutations in the key gene GJB2 of in nonsyndromic deafness（NSHI）are single nucleotide deletions and point mutations, resulting in mistranslation and non-function of Cx26 gap junction proteins. However, except for a few GJB2 mutations, there is no significant correlation between mutants and auditory phenotypes, and there is no clear relationship between mutant functions and phenotypes. In order to solve the problem that the traditional exome sequencing can not meet the requirements of in-depth study of its pathogenesis, our group uses transcriptome, translation group and apparent group sequencing data from Cx26-NSHI patients with different types of GJB2 gene mutations to integrate and analyze the linear or non-linear relationship between them, so as to effectively remove the random events of a single link. To study the molecular mechanism between GJB2 gene mutations and clinical phenotypes from a new perspective. At the same time, in order to repair the Cx26 defect caused by GJB2 gene mutation, our team constructed the transgenic mouse model of GJB2 gene defect, and used Gesicle-mediated gene editing technology to correct the mutation to restore or improve its function. Through the GJB2 gene mutation caused by the upstream Cx26-NSHI gene detection of disease modeling and then downstream gene therapy, to form an overall research idea.