CRISPR/Cas9 gene editing system and application of CRISPR/Cas9 gene editing system in preparation of medicine for treating genetic sensorineural deafness

Abstract

The invention discloses a CRISPR / Cas9 gene editing system and application of the CRISPR / Cas9 gene editing system in preparation of a medicine for treating genetic sensorineural deafness. The gene editing system comprises gRNA of a specific targeting Myo6C442Y mutant gene and SaCas9-KKH, and an effective knockout system aiming at Myo6C442Y is formed through AAV-PHP.eB virus vector delivery. According to the application disclosed by the invention, the influence of the AAV-SaCas9-KKH-Myo6-g2 system on the hearing loss of the mouse is evaluated through a Myo6WT / C442Y mouse model experiment, and the result shows that the Myo6C442Y mutant allele in the Myo6WT / C442Y mouse can be specifically knocked out by the AAV-SaCas9-KKH-Myo6-g2 system, the recovery of an auditory function can be observed within 5 months after the AAV-SaCas9-KKH-Myo6-g2 is injected into the Myo6WT / C442Y mouse, and meanwhile, the situation that the incubation period of ABR I wave is shortened, the cell survival rate is improved and the cilia form is regular are also observed; the system visually verifies the treatment effect of the AAV-SaCas9-KKH-Myo6-g2 system on the semi-dominant hereditary sensorineural deafness caused by the Myo6C442Y mutant gene, and can be used for preparing the medicine for treating the semi-dominant hereditary sensorineural deafness.

Description

technical field [0001] The invention relates to the fields of molecular biology and medicine, in particular to a CRISPR / Cas9 gene editing system and its application in the preparation of drugs for treating hereditary sensorineural deafness. Background technique [0002] Myosin VI (Myosin VI, MYO VI) is an unconventional myosin that is expressed in both the inner and outer hair cells of the inner ear, especially the epidermal plate of the cilium. important role. Pathogenic variants in MYO6 cause either autosomal dominant nonsyndromic hearing loss or autosomal recessive nonsyndromic hearing loss. A large-scale genetic analysis of hearing loss patients in Japan found that 2.4% of autosomal dominant sensorineural deafness cases were caused by mutations in MYO6. The myosin VI p.C442Y mutation causes progressive hearing loss in humans that begins in childhood and manifests as severe sensorineural deafness by age 5. Although MYO6 mutations play an important role in hereditary de...

AI Technical Summary

Problems solved by technology

[0005] The present invention targets deficiency therapy Myo6 C442Y The problem of deafness caused by mutated genes, the purpose is to provide a CRISPR / Cas9 gene editing system to specifically target and knock out Myo6 C442Y Mutating the gene, retaining the wild-type strand makes Myo6 WT Genes function normally, and thus treatment by Myo6 C442Y Semi-dominant hereditary sensorineural deafness caused by mutated genes can be used to prepare drugs for the treatment of hereditary sensorineural deafness

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