Electropermeabilization of skeletal muscle enhances gene transfer in vivo.
This work demonstrates that electrical muscle stimulation markedly increases the transfection efficiency of an intramuscular injection of plasmid DNA. In soleus or extensor digitorum longus muscles of adult rats the percentage of transfected fibers increased from about 1 to more than 10. The number of transfected fibers and the amount of foreign protein produced could be graded by varying the number or duration of the electrical pulses applied to the muscle. The stimulation had to be applied when DNA was present in the muscle. When dextran was injected together with the plasmid DNA, it was also taken up by the transfected fibers. Stimulation-induced membrane permeabilization and increased DNA uptake were therefore probably responsible for the improved transfection. The stimulation caused some muscle damage but the fibers regenerated rapidly. The described method, which is simple, efficient, and reproducible, should become valuable for basic research, gene therapy and DNA vaccination.
Mathiesen
I
I
Department of Physiology, University of Oslo, Norway.
D000818:Animals; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D008297:Male; D018482:Muscle, Skeletal / Q000378:metabolism*; D010957:Plasmids; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection / Q000379:methods*; D019444:Vaccines, DNA / Q000008:administration & dosage; D001616:beta-Galactosidase / Q000235:genetics
D019444:Vaccines, DNA; D001616:beta-Galactosidase