Optimisation of electrotransfer of plasmid into skeletal muscle by pretreatment with hyaluronidase -- increased expression with reduced muscle damage.
The efficiency of plasmid gene transfer to skeletal muscle can be significantly improved by the application of an electrical field to the muscle following injection of plasmid DNA. However, this electrotransfer is associated with significant muscle damage which may result in substantial loss of transfected muscle fibres. Reduction of the voltage used in the technique can result in a decrease in muscle damage, with a concomitant reduction in expression, but without a significant decrease in the number of transfected fibres. Pre-treatment of the muscle with a solution of bovine hyaluronidase greatly increases the efficiency of plasmid gene transfer when used in conjunction with electrotransfer, but not when used alone. This combination treatment results in greatly enhanced levels of transfected muscle fibres without the increases in muscle damage associated with the electrotransfer process.
McMahon
J M
JM
Gene Targeting Unit, Department of Neuromuscular Diseases, Division of Neuroscience and Psychological Medicine, Imperial College School of Medicine, Charing Cross Hospital, London, UK.
Signori
E
E
Wells
K E
KE
Fazio
V M
VM
Wells
D J
DJ
D000818:Animals; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D006821:Hyaluronoglucosaminidase / Q000008:administration & dosage*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D018482:Muscle, Skeletal / Q000201:enzymology* / Q000473:pathology; D009136:Muscular Dystrophies / Q000628:therapy*; D010957:Plasmids / Q000008:administration & dosage*; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D001616:beta-Galactosidase; D006821:Hyaluronoglucosaminidase