Figure 7
1746c27 delivery of a PMO therapeutic in vivo in a disease model of DMD. (a) Intracellular delivery of 1746c27_M23D(+7–18) induces dose-dependent skipping of exon 23 of the dystrophin gene in differentiated murine H-2Kb-tsA58 myogenic cultures. Exon skipping (marked by arrow) can be detected by RT-PCR from doses of 50 nM 1746c27_M23D(+7–18), but is not detected at any dose of M23D(+7–18) morpholino alone or the untreated cells (UT). (b) Tissue staining for dystrophin expression shows in vivo treatment of C57BL/10ScSnmdx mice (5 treatments over two weeks, with 4 nmol per dose) of 1746c27_M23D(+7–18) causes improved dystrophin protein levels and muscle architecture in the diaphragm, and to a lesser extent the tibialis anterior (samples taken from two independently-treated mice, n = 2). This improvement is compared to untreated C57BL/10ScSnmdx mice (Mdx untreated control) or those treated with the M23D(+7–18) morpholino alone (M23D(+7–18)-PMO). Treatment with Pip6-morpholino conjugate (Pip6_M23D(+7–18)-PMO) was a positive control for antisense-induced dystrophin expression. Tissue staining for dystrophin expression in C57BL/10ScSn mice (C57 untreated control) shows normal muscle architecture for comparison. Bar scale is 100 µm.
