Methodology article

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

Jordan Taylor¹ , Charlie F Babbs¹ , Mohammed Borhan Alzghoul² , Aaron Olsen¹ , Mickey Latour³ , Amber L Pond¹ and Kevin Hannon¹

¹  Department of Basic Medical Sciences, Purdue University, W. Lafayette IN 47907, USA

²  Department of Basic Veterinary Medical Science, Jordan University of Science and Technology, Irbid, Jordan

³  Department of Animal Sciences, Purdue University, W. Lafayette IN 47907, USA

author email corresponding author email

BMC Biotechnology 2004, 4:11doi:10.1186/1472-6750-4-11

Published:	18 May 2004

Abstract

Background

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.

Results

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 electroporation in vivo.

Conclusions

When injecting DNA intramuscularly, a flat plate electrode without any plasmid enhancers is the best method to achieve high levels of gene expression.