Kinetic of transgene expression after electrotransfer into skeletal muscle: importance of promoter origin/strength.
We determined over a 3-week period some of the factors that may influence the kinetic of gene expression following in vivo gene electrotransfer. Histochemical analysis of beta-galactosidase and biochemical analysis of luciferase expressions were used to determine reporter gene activity in the Tibialis anterior muscles of young Sprague-Dawley male rats. Transfection efficiency peaked 5 days after gene electrotransfer and then exponentially decreased to reach non-detectable levels at day 28. Reduction of muscle damage by decreasing the amount of DNA injected or the cumulated pulse duration did not improve the kinetic of gene expression. Electrotransfer of luciferase expression plasmids driven either by viral or mammalian promoters rather show that most of the decrease in transgene expression was related to promoter origin/strength. By regulating the amount of transgene expression, the promoter origin/strength could modulate the immune response triggered against the foreign protein and ultimately the kinetic of transgene expression.
Durieux
Anne-Cécile
AC
Laboratoire de Physiologie, Unité Physiologie et Physiopathologie de l'Exercice et Handicap, Université Jean Monnet, Faculté de Médecine, 42023 Saint-Etienne, France.
Bonnefoy
Régis
R
Freyssenet
Damien
D
D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000008:administration & dosage; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D007700:Kinetics; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019076:Transgenes / Q000502:physiology*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D004247:DNA; D008156:Luciferases; D001616:beta-Galactosidase