This study demonstrates that using a hair cell-specific Myo15 promoter in adeno-associated viral (AAV) gene therapy can effectively rescue hearing in a mouse model of autosomal recessive deafness (DFNB9). The Myo15 promoter was found to efficiently express the GFP reporter gene in hair cells while minimizing expression in other inner ear and central nervous system (CNS) cells. When used to deliver the OTOF gene, which is mutated in DFNB9, the Myo15 promoter significantly improved the exocytosis function of inner hair cells (IHCs) and restored hearing in Otof-/- mice. The study also showed that the Myo15 promoter allows for precise and efficient transgene expression in hair cells, reducing off-target effects and enhancing the safety of gene therapy. The results suggest that the Myo15 promoter could be a promising tool for treating hereditary hearing loss, particularly in cases involving hair cell dysfunction. The study highlights the importance of using cell-specific promoters to improve the efficacy and safety of gene therapy for hearing loss. The findings indicate that gene therapy mediated by the Myo15 promoter has potential for clinical application in treating autosomal recessive deafness and other hereditary hearing loss conditions.This study demonstrates that using a hair cell-specific Myo15 promoter in adeno-associated viral (AAV) gene therapy can effectively rescue hearing in a mouse model of autosomal recessive deafness (DFNB9). The Myo15 promoter was found to efficiently express the GFP reporter gene in hair cells while minimizing expression in other inner ear and central nervous system (CNS) cells. When used to deliver the OTOF gene, which is mutated in DFNB9, the Myo15 promoter significantly improved the exocytosis function of inner hair cells (IHCs) and restored hearing in Otof-/- mice. The study also showed that the Myo15 promoter allows for precise and efficient transgene expression in hair cells, reducing off-target effects and enhancing the safety of gene therapy. The results suggest that the Myo15 promoter could be a promising tool for treating hereditary hearing loss, particularly in cases involving hair cell dysfunction. The study highlights the importance of using cell-specific promoters to improve the efficacy and safety of gene therapy for hearing loss. The findings indicate that gene therapy mediated by the Myo15 promoter has potential for clinical application in treating autosomal recessive deafness and other hereditary hearing loss conditions.