Exon Exchange Approach to Repair Duchenne Dystrophin Transcripts
Figure 6
Exon 23 correction by exon replacement.
(A) The ExCh AS2-E23-DISE-AS4 molecule comprises the same elements as AS2-E24-DISE-AS4, except E23 instead of E24. E23 is the wild-type murine dystrophin E23 carrying a HindIII restriction site absent in the mdx minigene E23. (B) Expected transcripts generated by cis-splicing and E23 exchange, and predicted size of the corresponding RT-PCR products using primers A and C. (C) RT-PCR analysis using primers A and C of NIH3T3 cells cotransfected with dystrophin minigene and constructions pSMD2 (Ctrl) and ExCh plasmid pSMD2-AS2-E23-DISE-AS4; H20: PCR negative control. Representative results from two independent transfection experiments. (D) Cloning results of the RT-PCR amplicons obtained in (C) with AS2-E23-DISE-AS4 molecule. The clone numbers corresponding to parental and repaired inserts are described.