Electrotransfer of the full-length dog dystrophin into mouse and dystrophic dog muscles.
Duchenne muscular dystrophy (DMD) is an X-linked genetic disease characterized by the absence of dystrophin (427 kDa). An approach to eventually restore this protein in patients with DMD is to introduce into their muscles a plasmid encoding dystrophin cDNA. Because the phenotype of the dystrophic dog is closer to the human phenotype than is the mdx mouse phenotype, we have studied the electrotransfer of a plasmid carrying the full-length dog dystrophin (FLDYS(dog)) in dystrophic dog muscle. To achieve this nonviral delivery, the FLDYS(dog) cDNA was cloned in two plasmids containing either a cytomegalovirus or a muscle creatine kinase promoter. In both cases, our results showed that the electrotransfer of these large plasmids (∼17 kb) into mouse muscle allowed FLDYS(dog) expression in the treated muscle. The electrotransfer of pCMV.FLDYS(dog) in a dystrophic dog muscle also led to the expression of dystrophin. In conclusion, introduction of the full-length dog dystrophin cDNA by electrotransfer into dystrophic dog muscle is a potential approach to restore dystrophin in patients with DMD. However, the electrotransfer procedure should be improved before applying it to humans.
Pichavant
Christophe
C
Unité de Recherche en Génétique Humaine, Centre de Recherche du Centre Hospitalier de l'Université Laval, Centre Hospitalier Universitaire de Québec, and Faculté de Médecine, Université Laval, Sainte-Foy, Québec, Canada.
Chapdelaine
Pierre
P
Cerri
Daniel G
DG
Bizario
Joao C S
JC
Tremblay
Jacques P
JP
D000818:Animals; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D016189:Dystrophin / Q000235:genetics* / Q000378:metabolism*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D018101:Mice, Inbred mdx; D018345:Mice, Knockout; D018482:Muscle, Skeletal / Q000378:metabolism*; D009137:Muscular Dystrophy, Animal / Q000235:genetics / Q000378:metabolism*; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D014162:Transfection
D018076:DNA, Complementary; D016189:Dystrophin