Antisense PMO Found in Dystrophic Dog Model Was Effective in Cells from Exon 7-Deleted DMD Patient
Figure 4
Multi exon skipping and recovery of dystrophin in CXMDJ and DMD 8772-derived cells.
(A) Schematic diagram of the three- and four-antisense PMO cocktails. For DMD 8772, Ex6B was replaced with hEx6B. In the four-antisense PMO cocktail, one additional sequence (Ex8G, Ex8I, or Ex8K) was added to the three-antisense PMO cocktail. (B) RT-PCR of dystrophin mRNA isolated from MyoD-transduced fibroblasts after treatment with the three- and four-antisense PMO cocktails. In-frame exon skipping products were 99 bp in dog and 221 bp and 92 bp in human. (C) Representative immunostaining and immunoblotting analysis of MyoD-transduced fibroblasts treated with antisense PMO cocktails. The nuclei were counterstained with DAPI. Scale bar: 100 µm. Expected molecular weights of truncated human dystrophin with exons 6–8 and exons 6–9 skipped are 18.3 kDa and 23.1 kDa, respectively, smaller than the full-length dystrophin.