title: Muscle transfection by electroporation with high-voltage and short-pulse currents provides high-level and long-lasting gene expression.
abstract: 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.
authors.affiliation: INSERM U 318, Laboratoire de Neurobiophysique-UJFG, Centre Hospitalier Universitaire, Grenoble, France. Jean-Michel.Vicat@ujf-grenoble.fr
authors.forename: J M
authors.initials: JM
authors.lastname: Vicat
authors.forename: S
authors.initials: S
authors.lastname: Boisseau
authors.forename: P
authors.initials: P
authors.lastname: Jourdes
authors.forename: M
authors.initials: M
authors.lastname: Lainé
authors.forename: D
authors.initials: D
authors.lastname: Wion
authors.forename: R
authors.initials: R
authors.lastname: Bouali-Benazzouz
authors.forename: A L
authors.initials: AL
authors.lastname: Benabid
authors.forename: F
authors.initials: F
authors.lastname: Berger
mesh_terms: 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
chemical_list: D008156:Luciferases; D001616:beta-Galactosidase