新生儿中遗传性耳聋发病率高达千分之一，但尚无有效病因治疗。前期研究证实CRISPR/Cas9基因编辑可以治疗新生小鼠显性遗传性耳聋，CRISPR/Cas9精准治疗其他类型的遗传性耳聋及成年小鼠仍是空白。小鼠Pmca2基因点突变导致听力渐进性丧失和毛细胞进行性凋亡，具有单倍体显性不足表征，是利用CRISPR/Cas9治疗的良好模型。本研究利用CRISPR/Cas9对Pmca2突变新生小鼠（耳蜗微注射）和成年小鼠（后半规管注射）致聋基因进行修改，通过听性脑干诱发电位（ABR）／畸变产物耳声发射（DPOAE）检测听力，行为学评估听觉，免疫组化、电生理、双光子检测细胞功能，高通量测序检测基因编辑效率和脱靶率，以期证实CRISPR／Cas9基因编辑能够安全有效治疗新生和成年单倍体显性不足遗传性耳聋小鼠模型，并建立耳聋基因编辑方法学，为遗传性耳聋精准基因治疗临床转化提供依据。

Genetic hearing loss affects one in thousand newborns without available biological treatment. Previous study proved that CRISPR/Cas9 mediated gene editing can restore hearing of dominant genetic deafness mice, but whether gene editing could rescue other types of genetic deafness or adult mice is unclear. Pmca2 point mutant which causes progressive hearing loss and hair cell death in mice, showing aploinsufficiency deafness, is an ideal mouse model for gene therapy by CRISPR/Cas9. The purpose of this project is to modify mutation gene in newborn (Scala media micro-injection) and adult (Posterior semicircular canal injection) mouse, measuring hearing by Auditory Brainstem Response and Distortion Product Otoacoustic Emission, auditory behavior test, testing function in cell level by immunohistochemistry, electrophysiology and two-photon microscope, detecting gene editing efficiency and off-targets by next generation sequencing. We attempt to testify the possibility of gene therapy in neonatal and adult haploinsufficiency genetic deafness mice by CRISPR/Cas9, providing evidence for clinical trial of gene therapy on genetic deafness.