Abstract
Electroporation generally refers to the technique of permeabilizing cell membranes by applying a short and intense electric pulse across a cell, such that the barrier function of the membrane is instantaneously compromised. During such time, genetic materials may travel across the membrane. For a successful gene transfer process, the barrier function of the cell membrane is rapidly restored, and the cell survives. The electrotransfection process thus comprises two steps. The first step is electroporation, which is governed by the electrical properties of the cell and the suspension medium. The controlling parameters are mainly electrical. The second step is recovery, which must take into account the biological characteristics of the cells. We consider these two steps in this chapter.
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References
Neumann, E. and Rosenheck, K. (1972) Permeability changes induced by electric impulses in vesicular membranes,J. Membr. Biol. 10, 279–290.
Zimmermann, U., Pilwat, G., and Riemann, F. (1974) Dielectric breakdown of cell membranes. Biophys. J. 14, 881–899.
Kinosita, K. and Tsong, T. Y. (1975) Formation and reealing of pores of controlled sizes in human erythrocyte membranes. Nature 268, 438–441.
Zimmermann, U., Riemann, F., and Pilwat, G. (1976) Enzyme loading of electrically homogeneous human red blood cell ghosts prepared by dielectric breakdown. Biochim. Biophys. Acta 436, 460–474.
Vienken, J., Jeltsch, E., and Zimmermann, U. (1978) Penetration and entrapment of large particles in erythrocytes by electrical breakdown techniques. Cytobiologie 17, 182–196.
Auer, D., Brandner, G., and Bodemer, W. (1976) Dielectric breakdown of the red blood cell membrane and uptake of SV40 DNA and mammalian RNA. Naturwissenschaften 63, 391–393.
Wong, T. K. and Neumann, E. (1982) Electric field mediated gene transfer. Biochem. Biophys. Res. Commun. 107, 584–587.
Xie, T. D., Sun, L., and Tsong, T. Y. (1990) Study of mechanisms of electrid field-induced DNA transfection, I. DNA entry by surface binding and diffusion through membrane pores. Biophys. J. 58, 13–19.
Abidor, I. G., Arakelyan, V. B., Chernomordik, L. V., and Chizmadzhev, Yu., A. (1979) Electric breakdown of bilayer membranes. I. The main experimental facts and their qualitative discussion. Bioelectrochem. Bioenerg. 6, 37–52.
Zelev, D. V. and Needham, D. (1993) Tension-stabilized pores in gaint vesicles: Determination of pore size and pore line tension. Biochim. Biophys. Acta 1147, 89–104.
Holzapfel, C. J., Vienkan, J., and Zimmermann, U. (1982) Rotation of cells in an alternating electric field: Theory and experimental proof. J. Membr. Biol. 67, 13–26.
Rols, M. P. and Teissié, J. (1990) Electropermeabilization of mammalian cells: Quantitative analysis of the phenomenon. Biophys. J. 58, 1089–1098.
Stenger, D. A., Kaler, K. V. I. S., and Hui, S. W. (1991) Dipole interactions in electrofusion: contributions of membrane potential and effective dipole to interaction pressure. Biophys. J. 59, 1074–1084.
Needham, D. and Hochmuth, R. M. (1989) Electro-mechanical permeabilization of lipid vesicles: Role of membrane tension and compressibility. Biophys. J. 55, 1001–1009.
Klenchin, V. A., Sukharev, S. I., Serov, S. M., Chernomodok. L. V., and Chizmadzhev, Yu. A. (1991) Electrically induced DNA uptake by cells is a fast process involving DNA electrophoresis. Biophys. J. 60, 804–811.
Wolf, H., Rols, M. P., Boldt, E. Neumann, E., and Teissié, J. (1994) Control by pulse parameters of electric field-mediated gene transfer in mammalian cells. Biophys. J. 66, 524–531.
Andreason, G. L. and Evans, G. A. (1989) Optimization of electroporation for transfection of mammalian cells. Anal. Biochem. 180, 269–275.
Sukharev, S. I., Klenchin, V. A., Serov, S. M., Chernomodok. L. V., and Chizmadzhev, Yu. A. (1992) Electroporation and electrophoretic DNA transfer into cells: The effect of DNA interaction with electropores. Biophys. J. 63, 1320–1327.
Nickoloff, J. A. and Reynolds, R. J. (1992) Electroporation-mediated gene transfer efficiency is reduced by linear plasmid carrier DNAs. Anal. Biochem. 205, 237–243.
Hibino, M., Shigemori, M., Itoh, H., Nagayama, K., and Kinosita, K. Jr. (1991) Membrane conductance of an electroporated cell analyzed by submicrosecond imaging of transmembrane conductance. Biophys. J. 59, 209–220.
Schwister, K. and Deuticke, B. (1985) Formation and properties of aqueous leaks induced in human erythrocytes by electrical breakdown. Biochim. Biophys. Acta 816, 332–348.
Liang, H., Purucker, W. J., Stenger, D. A., Kubiniec, R. T., and Hui, S. W. (1988) Uptake of fluorescence-labeled dextrans by 10T-1/2 fibroblasts following permeation by rectangular and exponential-decay electric field pulse. Biotechniques 6, 550–558.
Prausnitz, M. R., Milano, C. D., Gimm, J. A., Langer, R., and Weaver, J. C. (1994) Quantitative study of molecular transport due to electroporation: Uptake of bovine serum albumin by erythrocyte ghosts. Biophys. J. 66, 1522–1530.
Kubiniec, R. T., Liang, H., and Hui, S. W. (1990) Effects of pulse length and pulse strength on transfection by electroporation. Biotechniques 8, 1–3.
Xie, T. D. and Tsong, T. Y. (1992) Study of mechanisms of electrid field-induced DNA transfection, III. Electric parameters and other conditions for effective transfection. Biophys. J. 63, 28–34.
Chang, D. C., Chassy, B. M., Saunders, J. A., and Sowers, A. E. (1992) Guide to Electroporation and Electrofusion. Academic Press, San Diego, CA.
Baker, P. F. and Knight, D. E. (1983). High voltage techniques for gaining access to the interior of cells: Application to the study of exocytosis and membrane turnover. Methods Enzymol. 98, 23–37.
Abidor, I. G., Li, L. H., and Hui, S. W. (1994) Studies of cell pellets. II. Osmotic properties, electroporation and related phenomena. Membrane interactions. Biophys. J. 67, 427–435.
Li, L. H., Ross, P., and Hui, S. W. (1999) Improving electrotransfection efficiency by post-pulse centrifugation. Gene Ther. 6, 364–372.
Hui, S. W., Stoicheva, N., and Zhao, Y-L. (1996) High efficiency loading, transfection and fusion of cells by electroporation in two-phase polymer systems. Biophys. J. 71, 1123–1130.
Li, L. H., Sen, A., Murphy, S. P., Jahreis, G. P., and Hui, S. W. (1999) DNAupdate induces apoptosis and limits transfection efficiency. Exp. Cell Res. (in press).
Chang, D. C., Gao, P. Q., and Maxwell, B. L. (1991) High efficiency gene transfection by electroporation using a radio-frequency electric field. Biochim. Biophys. Acta 1992, 153–160.
Tekle, E., Austumian, R. D., and Chock, P. B. (1991) Electroporation by using bipolar oscillating electric field: an improved method for DNA trandfection of NIH/3T3 cells. Proc. Natl. Acad. Sci. USA 88, 4230–4234.
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© 2000 Humana Press Inc., Totowa, NJ
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Hui, S.W., Li, L.H. (2000). In Vitro and Ex Vivo Gene Delivery to Cells by Electroporation. In: Jaroszeski, M.J., Heller, R., Gilbert, R. (eds) Electrochemotherapy, Electrogenetherapy, and Transdermal Drug Delivery. Methods in Molecular Medicine, vol 37. Humana Press. https://doi.org/10.1385/1-59259-080-2:157
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DOI: https://doi.org/10.1385/1-59259-080-2:157
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