Fig. 3

Restoration of dystrophin expression by CRISPR/Cas9. a CRISPR/Cas9-mediated gene editing restored dystrophin mRNA level in myotubes differentiated from DMD–MDSCs. Illustration of primer binding sites (black arrow) for RT-PCR (top row); all detected bands were of the expected size; red arrows indicate bands that were verified by Sanger sequencing in b. M, marker; Un, unedited. b The successful reframing of cDNA in the DMD–MDSCs subjected to three different gene-editing strategies was confirmed by Sanger sequencing. c Western blot analysis of dystrophin expression in targeted (∆45–55, ∆46–54, and INDEL50) or untargeted myotubes differentiated from the DMD–MDSCs; WT MDSCs served as a positive control. Expected molecular weights were 427 kDa for WT and INDEL50, 361 kDa for ∆45–55, and 375 kDa for ∆46–54; MHC served as a loading control. d Representative images of MHC (green) and dystrophin (red, white arrow) expression in targeted (∆45–55, ∆46–54, and INDEL50) and untargeted myotubes differentiated from the DMD–MDSCs as determined by immunocytochemistry; WT MDSCs served as a positive control, and nuclei were stained with DAPI (blue). Scale bar, 100 μm