1. Real time electroporation control for accurate and safe in vivo non-viral gene therapy
- Author
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Lluis M. Mir, Damijan Miklavčič, D. Cukjati, Franck M. Andre, Danute Batiuskaite, Vectorologie et transfert de gènes (VTG / UMR8121), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Faculty of Electrical Engineering, University of Ljubljana, Faculty of Natural Sciences - Department of Biology, and Vytautas Magnus University - Vytauto Didziojo Universitetas (VDU)
- Subjects
Electrochemotherapy ,MESH: Microelectrodes ,Genetic enhancement ,Cell ,MESH: Equipment Failure Analysis ,0302 clinical medicine ,Electrochemistry ,MESH: Animals ,0303 health sciences ,MESH: Muscle, Skeletal ,MESH: Feedback ,Electroporation ,MESH: DNA ,General Medicine ,Transfection ,Equipment Design ,Cell biology ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM] ,medicine.anatomical_structure ,Liver ,Naked DNA ,030220 oncology & carcinogenesis ,Viruses ,Female ,MESH: Rats ,Biophysics ,Biology ,Sensitivity and Specificity ,Feedback ,MESH: Viruses ,03 medical and health sciences ,In vivo ,Computer Systems ,medicine ,Animals ,Luciferase ,MESH: Electroporation ,Physical and Theoretical Chemistry ,Rats, Wistar ,Muscle, Skeletal ,030304 developmental biology ,MESH: Transfection ,MESH: Computer Systems ,MESH: Rats, Wistar ,DNA ,Genetic Therapy ,Molecular biology ,MESH: Sensitivity and Specificity ,Rats ,Equipment Failure Analysis ,MESH: Gene Therapy ,MESH: Female ,Microelectrodes ,MESH: Equipment Design ,MESH: Liver - Abstract
In vivo cell electroporation is the basis of DNA electrotransfer, an efficient method for non-viral gene therapy using naked DNA. The electric pulses have two roles, to permeabilize the target cell plasma membrane and to transport the DNA towards or across the permeabilized membrane by electrophoresis. For efficient electrotransfer, reversible undamaging target cell permeabilization is mandatory. We report the possibility to monitor in vivo cell electroporation during pulse delivery, and to adjust the electric field strength on real time, within a few microseconds after the beginning of the pulse, to ensure efficacy and safety of the procedure. A control algorithm was elaborated, implemented in a prototype device and tested in luciferase gene electrotransfer to mice muscles. Controlled pulses resulted in protection of the tissue and high levels of luciferase in gene transfer experiments where uncorrected excessive applied voltages lead to intense muscle damage and consecutive loss of luciferase gene expression.
- Published
- 2006
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