利用CRISPR-Cas9技术研究POU4F3基因在斑马鱼听觉系统中的功能及机制

耳聋是最普遍的感官缺陷疾病之一，60%以上的感音神经性聋由遗传因素导致。我们前期收集了一个常染色体显性遗传的迟发性非综合征型耳聋家系，利用耳聋基因目标区域捕获及大规模平行测序，发现一个POU4F3基因的新突变c. 337C>T（p.Gln113*），该突变导致了POU4F3蛋白的截短。然而，POU4F3基因的致聋机制还未完全揭示。斑马鱼是研究听觉疾病的理想模式生物，POU4F3在斑马鱼听觉系统的功能尚未系统研究。本课题拟利用最新的基因编辑技术——CRISPR-Cas9技术构建斑马鱼POU4F3的突变体模型。在成功建模的基础上，研究POU4F3在斑马鱼听觉系统中的时空表达定位；分析POU4F3在听觉系统的功能；进而在分子、细胞和动物水平开展系统的研究，从不同层次揭示POU4F3基因突变致聋的分子机制。本研究在遗传性耳聋的病因诊断、预防干预、新药研发和基因治疗等方面均具有重要价值。

Hearing loss is one of the most common sensory disorders, and over 60% of hearing loss patients can be attributed to genetic causes. Taking advantage of targeted capture and massive sequence of deafness genes, we identified a novel mutation c. 337C>T in POU4F3 gene from an autosomal dominantly non-syndromic sensorineural hearing loss family, which leading to a truncated protein. However, the pathogenic mechanism is not fully found yet. We aim to construct the POU4F3 mutant zebrafish model, to reveal the expression of POU4F3 and its function in auditory system of zebrafish. Based on the preliminary study, the present project is designed to investigate that: 1) the localization of POU4F3 at different development periods, 2) the role of POU4F3 in auditory system, 3) the pathogenic mechanism underlying POU4F3 gene related hearing loss. This research will lay the foundation of effective molecular diagnosis, intervention, development of drugs and genetic treatment measure for hereditary deafness.