Antisense oligonucleotide induced exon skipping has emerged as a promising therapeutic strategy for Duchenne muscular dystrophy (DMD). Systemic administration of antisense phosphorodiamidate morpholino oligomers (PMOs) targeting exons 6 and 8 in dystrophin mRNA of the canine X-linked muscular dystrophy model in Japan (CXMDJ) that lacks exon 7, restored dystrophin expression throughout skeletal muscle and ameliorated skeletal muscle pathology and function. However, the antisense PMO regime used in CXMDJ could not be considered for a direct application to DMD patients so far, because this type of mutation is quite rare. The authors have identified a DMD patient with an exon 7 deletion; and tried a direct translation of the antisense PMOs used in dog models to the DMD patient’s cells. This paper summarises the methods and protocols of multi-exon skipping using antisense PMOs in cells obtained from this DMD patient and CXMDJ dogs after performing MyoD-transduction and flow cytometry sorting.
