Optimization of ectopic gene expression in skeletal muscle through DNA transfer by electroporation

• Main Authors: Latour Mickey, Olsen Aaron, Alzghoul Mohammed, Babbs Charlie F, Taylor Jordan, Pond Amber L, Hannon Kevin
• Format: Article
• Language: English
• Published: BMC 2004-05-01
• Series: BMC Biotechnology
• Online Access: http://www.biomedcentral.com/1472-6750/4/11

<p>Abstract</p> <p>Background</p> <p>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 <it>in vivo</it>. Specifically, we investigated the effects of varying injection techniques, electrode surface geometry, and plasmid mediums.</p> <p>Results</p> <p>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 <it>in vitro</it>, had no positive effect on DNA electroporation <it>in vivo</it>.</p> <p>Conclusions</p> <p>When injecting DNA intramuscularly, a flat plate electrode without any plasmid enhancers is the best method to achieve high levels of gene expression.</p>