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Gene electrotransfer: Wikis


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Gene electrotransfer is a versatile biotechnology technique that enables transfer of genetic material into prokaryotic or eukaryotic cells. It is based on a physical method named electroporation, where transient increase in the permeability of cell membrane is achieved when submitted to short and intense electric pulses. In consequence transport of large molecules (naked plasmid DNA, antisense oligonucleotides, siRNA) into cells that otherwise cannot permeate through the cell membrane is achieved. Gene electrotransfer was first described in the 80s[1] and since then due to its ease of application and efficiency become a routine method for introducing foreign genes into bacterial,[2] yeast,[3] plant,[4] and animal cells [5] in vitro and into different tissues, including muscle,[6] tumors,[7] liver,[8] and skin[9] in vivo.


Physical principle

Application of electric pulses of sufficient strength to the cell causes an increase in the trans-membrane potential difference, which provokes the membrane destabilization. Cell membrane permeability is increased and otherwise nonpermeant molecules enter the cell.[10] Although the mechanisms of gene electrotransfer are not yet fully understood, it was shown that the introduction of DNA only occurs in the part of the membrane facing the cathode and that several steps are needed for successful transfection: electrophoretic migration of DNA towards the cell, DNA insertion into the membrane, translocation across the membrane, migration of DNA towards the nucleus, transfer of DNA across the nuclear envelope and finally gene expression.[11] There are a number of factors that can influence the efficiency of gene electrotransfer, such as: temperature, parameters of electric pulses, DNA concentration, electroporation buffer used, cell size and the ability of cells to express transfected genes.[12] In in vivo gene electrotransfer also DNA diffusion through extracellular matrix, properties of tissue and overall tissue conductivity are crucial.[13]

Biotechnological applications

Bacterial transformation is generally the easiest way to make large amounts of a particular protein needed for biotechnology purposes or in medicine. Since gene electrotransfer is very simple, rapid and highly effective technique it first became very convenient replacement for other transformation procedures.[14]

Medical applications

First medical application of electroporation was used for introducing poorly permeant anticancer drugs into tumor nodules.[15] Soon also gene electrotransfer became of special interest because of its low cost, easiness of realization and safety. Namely, viral vectors can have serious limitations in terms of immunogenicity and pathogenicity when used for DNA transfer.[16] Presently, gene electrotransfer is in preclinical phase and it is used to deliver large variety of therapeutic genes for treatment of several diseases, such as: disorders in immune system, tumors, organ or site-specific diseases, metabolic disorders, monogenetic diseases, cardiovascular diseases, analgesia….[17][18][19] Although the method is not systemic, but strictly local one, it is still the most efficient non-viral strategy for gene delivery.


  1. ^ Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH (1982). Gene transfer into mouse lyoma cells by electroporation in high electric fields, EMBO J 7:841-845
  2. ^ Drury L (1996). Transformation of bacteria by electroporation, Methods Mol Biol 58: 249-256
  3. ^ Simon JR (1993). Transformation of intact yeast cells by electroporation, Methods Enzymol 217:478-483
  4. ^ Terzaghi WB, Cashmore AR (1997). Plant cell transfection by electroporation, Methods Mol Biol 62:453-462
  5. ^ Kanduser M, Miklavcic D, Pavlin M (2009). Mechanisms involved in gene electrotransfer using high- and low-voltage pulses -An in vitro study, Bioelectrochemistry 74:265-271
  6. ^ Aihara H, Miyazaki J (1998). Gene transfer into muscle by electroporation in vivo, Nat Biotechnol 16:867-870
  7. ^ Cemazar M, Sersa G, Wilson J, Tozer GM, Hart SL, Grosel A, Dachs GU (2002). Effective gene transfer to solid tumors using different nonviral gene delivery techniques: Electroporation, liposomes, and integrin-targeted vector, Cancer Gene Ther 9:399-406
  8. ^ Heller R, Jaroszeski M, Atkin A, Moradpour D, Gilbert R, Wands J, Nicolau C (1996). In vivo gene electroinjection and expression in rat liver, FEBS Lett 389:225-228
  9. ^ Pavselj N, Preat V (2005). DNA electrotransfer into the skin using a combination of one high- and one low-voltage pulse, J Control. Release 106:407-415
  10. ^ Kotnik T, Miklavcic D (2000). Analytical description of transmembrane voltage induced by electric fields on spheroidal cells, Biophys J 79:670-679
  11. ^ Satkauskas S, Bureau MF, Puc M, Mahfoudi A, Scherman D, Miklavcic D, Mir LM (2002). Mechanisms of in vivo DNA electrotransfer: respective contributions of cell electropermeabilization and DNA electrophoresis, Mol Ther 5:133-140
  12. ^ Gehl J (2003). Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research, Acta Physiol Scand 177:437-447
  13. ^ Miklavcic D, Beravs K, Semrov D, Miklavcic D, Cemazar M, Demsar F, Sersa G (1998). The importance of electric field distribution for effective in vivo electroporation of tissues, Biophys J 74:2152–2158
  14. ^ Calvin NM, Hanawalt PC (1988). High-efficiency transformation of bacterial cells by electroporation, J Bacteriol 170:2796-801
  15. ^ Mir LM, Belehradek M, Domenge C, Orlowski S, Poddevin B, Belehradek J Jr, Schwaab G, Luboinski B, Paoletti C (1991). Electrochemotherapy, a new antitumor treatment: first clinical trial, CR Acad Sci III 313:613-618
  16. ^ Marshall E (1999). Gene therapy death prompts review of adenovirus vector, Science 286:2244–2245
  17. ^ Heller LC, Coppola D (2002). Electrically mediated delivery of vector plasmid DNA elicits an antitumor effect, Gene Ther 9:1321-1325
  18. ^ Chuang IC, Jhao CM, Yang CH, Chang HC, Wang CW, Lu CY, Chang YJ, Lin SH, Huang PL, Yang LC (2004). Intramuscular electroporation with the proopiomelanocortin gene in rat adjuvant arthritis, Arthritis Res Ther 6:R7–R14
  19. ^ Vilquin JT, Kennel PF, Paturneau-Jouas M, Chapdelaine P, Boissel N, Delaère P, Tremblay JP, Scherman D, Fiszman MY, Schwartz K (2001). Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies, Gene Ther 8:1097-1107


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