Precise detection of CRISPR-Cas9 editing in hair cells in the treatment of autosomal dominant hearing loss

Precise detection of CRISPR-Cas9 editing in hair cells in the treatment of autosomal dominant hearing loss

Blog Article

Gene therapy would benefit from the effective editing of targeted cells with CRISPR-Cas9 tools.However, it is difficult to precisely assess the editing performance in vivo because the tissues contain many non-targeted cells, which is one of the major barriers to clinical translation.Here, in the Atoh1-GFP;Kcnq4+/G229D mice, recapitulating a novel mutation we identified in a hereditary hearing loss pedigree, the high-efficiency editing Wire Loops of CRISPR-Cas9 in hair cells (34.10% on average) was precisely detected by sorting out labeled cells compared with only 1.

45% efficiency NUTRI FLEX in the whole cochlear tissue.After injection of the developed AAV_SaCas9-KKH_sgRNA agents, the Kcnq4+/G229D mice showed significantly lower auditory brainstem response (ABR) and distortion product otoacoustic emission (DPOAE) thresholds, shorter ABR wave I latencies, higher ABR wave I amplitudes, increased number of surviving outer hair cells (OHCs), and more hyperpolarized resting membrane potentials of OHCs.These findings provide an innovative approach to accurately assess the underestimated editing efficiency of CRISPR-Cas9 in vivo and offer a promising strategy for the treatment of KCNQ4-related deafness.