Electroporation of Plasmid DNA to Swine Muscle
For plasmid-mediated gene therapy applications, a major limitation to scale up from rodents to large animals is the low expression level of injected plasmid DNA. The electroporation technique, which results in the passage of foreign material through the cell membrane, is one method that has been shown to be effective at improving local plasmid uptake and consequently, expression levels. Previous studies have determined that optimized electroporation parameters (such as electric field intensity, number of pulses, lag time between plasmid injections and electroporations, and optimal plasmid formulation conditions) are dependent on the target muscle type and individual species. Here, we provide a detailed protocol to optimize conditions for the successful intramuscular electroporation of plasmid DNA to swine, a large animal model. Our results suggest that the technique is safe and effective for veterinary applications. Furthermore, these results provide evidence for the feasibility of upcoming human applications.
Key wordsElectroporation Gene therapy Muscle Plasmid DNA Swine
The authors would like to thank Dr. Patricia Brown for her help as an attending veterinarian to oversee the animal anesthesia and perform the electroporation animal procedures. This work was supported by Inovio Pharmaceuticals (Blue Bell, PA).
- 2.Draghia-Akli, R., Khan, A.S., Cummings, K.K., Parghi, D., Carpenter, R.H., Brown, P.A. (2002) Electrical enhancement of formulated plasmid delivery in animals. Tech Cancer Res Treat 1, 365–371.Google Scholar