Figure 5.
Formation of the Hybrid Exons 3-47/16-58 and 5-47/18-58 Restored the Expression of Dystrophin Protein In Vivo in Cardiomyocytes of the del52hDMD/mdx Mouse Model
4- to 5-week-old del52hDMD/mdx mice were systemically injected in the tail vein with a total amount of 7.5 × 1013 vg/kg of AAV9s coding for SaCas9 and for a pair of sgRNAs (i.e., either sgRNAs 3-47 and 16-58 or sgRNAs 5-47 and 18-58). Mice were euthanized 6 weeks after the injection, and tissues were collected. (A) Heart cryo-sections were stained with a rabbit polyclonal anti-dystrophin antibody (ab15277, Abcam). (B) PCR confirmed formation of the hybrid exons 47-58 in the hearts of treated mice. (C) The presence of the internally truncated dystrophin protein was confirmed by western blot analysis. (D) The amplicon sequences of the hybrid exons were submitted to TIDE analysis to determine the percentage of exact junction (n = 4). The error bars are SE.