Optimization of Ectopic Gene Expression in Skeletal Muscle Through DNA Transfer by Electroporation.
Electroporation (EP) is a widely used non-viral gene transfer method. We have attempted to develop an exact protocol to maximize DNA expression while minimizing tissue damage following EP of skeletal muscle in vivo. Specifically, we investigated the effects of varying injection techniques, electrode surface geometry, and plasmid mediums.
We found that as the amount of damage increased in skeletal muscle in response to EP, the level of β-galactosidase (β-gal) expression drastically decreased and that there was no evidence of β-gal expression in damaged fibers. Two specific types of electrodes yielded the greatest amount of expression. We also discovered that DNA uptake in skeletal muscle following intra-arterial injection of DNA was significantly enhanced by EP. Finally, we found that DMSO and LipoFECTAMINE™, common enhancers of DNA electroporation in vitro, had no positive effect on DNA electroporationin vivo.
When injecting DNA intramuscularly, a flat plate electrode without any plasmid enhancers is the best method to achieve high levels of gene expression.
Date of this Version
Taylor, Jordan; Babbs, Charles F.; Alzghoul, Mohammed Borhan; Olsen, Aaron; Latour, Mickey; Pond, Amber L.; and Hannon, Kevin, "Optimization of Ectopic Gene Expression in Skeletal Muscle Through DNA Transfer by Electroporation." (2004). Weldon School of Biomedical Engineering Faculty Publications. Paper 15.
This is the publisher pdf of Jordan Taylor, Charlie F Babbs, Mohammed Alzghoul, Aaron Olsen, Mickey Latour, Amber L Pond, Kevin Hannon. Optimization of Ectopic Gene Expression in Skeletal Muscle Through DNA Transfer by Electroporation. BMC Biotechnology 2004, 4:11 (18 May 2004) and is available at: 10.1186/1472-6750-4-11.