Prime Editing Enables High-Efficiency Correction of the Ryr1 T4706M Mutation: A Promising Therapeutic Approach for RyR1-Related Myopathies

Abstract

Prime editing has emerged as a powerful genome-editing tool for precise correction of pathogenic mutations, offering a promising therapeutic approach for genetic myopathies. Here, we evaluate the correction efficiency of the T4706M mutation in the Ryr1 gene, which is implicated in severe skeletal muscle dysfunction. Using an optimized epegRNA design and RNA electroporation, we achieved a remarkable 80% editing efficiency in immortalized C2C12 myoblasts and 37% correction in primary myoblasts derived from the RYR1TM/TM mouse model. Our results demonstrate that the PE6 prime editing strategy, combined with rationally designed epegRNAs, significantly enhances editing efficiency in unselected cell populations. These findings establish a critical ex vivo foundation for the development of in vivo Ryr1 gene therapy in preclinical mouse models. They also provide a validated editing design that can support delivery-focused applications in both academic and industry settings.