Muscle transfection by electroporation with high-voltage and short-pulse currents provides high-level and long-lasting gene expression.
Gene transfer into muscle by electroporation with low-voltage and long-pulse (LV/LP, 100 V/50 msec) currents was shown to be more efficient than simple intramuscular DNA injection. Nevertheless, transgene expression declined from day 7 and only reached 10% of the maximum 3 weeks after electroporation. We have optimized electroporation conditions including voltage, pulse number, and the amount of injected luciferase-encoding plasmid DNA in the tibialis anterior muscle. Using high-voltage and short-pulse (HV/SP, 900 V/100 microsec) currents, we observed an average 500-fold increase in luciferase expression, in comparison with nonelectroporated muscle. Moreover, sustained and long-lasting gene expression was observed for at least 6 months. When we compared HV/SP currents with LV/LP currents, luciferase expression was similar 24 hr after electroporation. One month later, whereas luciferase expression was stable in muscle electroporated with HV/SP currents, it decreased 600-fold in muscle electroporated with LV/LP currents. In conclusion, electroporation with high-voltage and short-pulse currents provides high-level and long-lasting gene expression in muscle.
Vicat
J M
JM
INSERM U 318, Laboratoire de Neurobiophysique-UJFG, Centre Hospitalier Universitaire, Grenoble, France. Jean-Michel.Vicat@ujf-grenoble.fr
Boisseau
S
S
Jourdes
P
P
Lainé
M
M
Wion
D
D
Bouali-Benazzouz
R
R
Benabid
A L
AL
Berger
F
F
D000818:Animals; D004553:Electric Conductivity; D018274:Electroporation / Q000379:methods*; D005260:Female; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018482:Muscle, Skeletal / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D019076:Transgenes; D001616:beta-Galactosidase / Q000378:metabolism
D008156:Luciferases; D001616:beta-Galactosidase