Plasmid DNA electrotransfer for intracellular and secreted proteins expression: new methodological developments and applications.
In vivo electrotransfer is a physical method of gene delivery in various tissues and organs, relying on the injection of a plasmid DNA followed by electric pulse delivery. The importance of the association between cell permeabilization and DNA electrophoresis for electrotransfer efficiency has been highlighted. In vivo electrotransfer is of special interest since it is the most efficient non-viral strategy of gene delivery and also because of its low cost, easiness of realization and safety. The potentiality of this technique can be further improved by optimizing plasmid biodistribution in the targeted organ, plasmid structure, and the design of the encoded protein. In particular, we found that plasmids of smaller size were electrotransferred more efficiently than large plasmids. It is also of importance to study and understand kinetic expression of the transgene, which can be very variable, depending on many factors including cellular localization of the protein, physiological activity and regulation. The most widely targeted tissue is skeletal muscle, because this strategy is not only promising for the treatment of muscle disorders, but also for the systemic secretion of therapeutic proteins. Vaccination and oncology gene therapy are also major fields of application of electrotransfer, whereas application to other organs such as liver, brain and cornea are expanding. Many published studies have shown that plasmid electrotransfer can lead to long-lasting therapeutic effects in various pathologies such as cancer, blood disorders, rheumatoid arthritis or muscle ischemia. DNA electrotransfer is also a powerful laboratory tool to study gene function in a given tissue.
Laboratoire de Pharmacologie Chimique et Génétique, U266 INSERM FRE 2463 CNRS, Faculté de Pharmacie Paris 5/ENSCP, 4 avenue de l'Observatoire, 75270 Paris cedex 06, France.
Carole
C
Bloquel
Emmanuelle
E
Fabre
Michel Francis
MF
Bureau
Daniel
D
Scherman
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D007700:Kinetics; D051379:Mice; D016513:Mice, SCID; D018482:Muscle, Skeletal / Q000502:physiology*; D010957:Plasmids; D019076:Transgenes