Gene repair in duchenne muscular dystrophy

Abstract

This study seeks to demonstrate gene repair in DMD gene deletion by exon insertion/replacement mediated by RNA splicing. A mini-gene target simulating a mutation model of exon 60 deletion of dystrophin was constructed. Six pre-therapeutic molecules (PTMs) consisting of a 5a?? and 3a?? binding domain, trans-splicing domain and WT replacement exons of 60 and 61 were artificially constructed.The results showed that WT exon 60 was inserted successfully and the trans-splicing efficiency ranged from 10% to 44% by semi quantitative assay. Furthermore, comparison between the six different designs of PTMs showed that masking of endogenous splicing elements of the target mini-gene transcript was critical. Variations in the length of the binding domains were less effective in inducing trans-splicing as compared to the role of the splicing elements. The findings from this study suggest that this approach of trans-splicing mediated exon insertion/replacement may provide a novel strategy for DMD gene repair.