Abstract
Introduction of foreign molecules, such as DNA, RNA, or proteins, into living cells is a powerful means to test the biological functions of these molecules. One of the techniques by which foreign molecules can be introduced into living cells is electroporation (for reviews, see refs. 1,2). Compared to other techniques for the introduction of molecules into living cells, electroporation does not seem to cause prolonged alterations in the biological structure and function of the target cells. Nevertheless, the pores that are induced by the electroporation technique (3–5) can be visualized by rapid-freezing electron microscopy techniques (6) and allow a direct contact between the cytoplasm of the cells and the electroporation medium. The components of the electroporation medium can, therefore, penetrate into the cell interior as long as the pores exist (7). In this perspective, it is obvious that cells are exquisitely sensitive to the composition of the electroporation medium (8–10) and that an electroporation medium that resembles the composition of the cytoplasm would probably enhance the viability of the cells after electroporation.
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References
Chang, D. C., Chassy, B. M., Saunders, J. A., and Sowers, A. E. (1992) Guide to Electroporation and Electrofusion. Academic, San Diego.
Alam, J., and Cook, J. C. (1990) Reporter genes: application to the study of mammalian gene transcription. Anal. Biochem. 188, 245–254.
Saulis, G., Venslauskas, M. S., and Naktinis, J. (1991) Kinetics of pore resealing in cell membranes after electroporation. Bioeletroch. Bioenerg. 26, 1–13.
Chernomordik, L. V., Sukharev, S. I., Popov, S. V., Pastushenko, V. F., Sokirko, A. V., Abidor, I. G., and Chizmadzhev, Y. A. (1987) The electrical breakdown of cell and lipid membranes: the similarity of phenomenologies. Biochim. Biophys. Acta. 902, 360–373.
Tsong, T. Y. (1991) Electroporation of cell membranes. Biophys. J. 60, 297–306.
Chang, D. C., and Reese, T. S. (1990) Changes in membrane structure induced by electroporation as revealed by rapid-freezing electron microscopy. Biophys. J. 58, 1–12.
Tekle, E., Astumian, R. D., and Chock, P. N. (1991) Electroporation by using bipolar oscillating electric field: an improved method for DNA transfection of NIH 3T3 cells. Proc. Natl. Acad. Sci. USA 88, 4230–4234.
Chu, G., Hayakawa, H., and Berg, P. (1987) Electroporation for the efficient transfection of mammalian cells with DNA. Nucleic Acids Res. 15, 1311–1326.
van den Hoff, M. J. B., Labruyère, W. T., Moorman, A. F. M., and Lamers, W. H. (1990) The osmolarity of the electroporation medium affects the transient expression of genes. Nucleic Acids Res. 18, 6464.
van den Hoff, M. J. B., Moorman, A. F. M., and Lamers, W. H. (1992) Electroporation in “intracellular” buffer increases cell survival. Nucleic Acids Res. 20, 2902.
Knight, D. E., and Scrutton, M. C. (1986) Gaining access to the cytosol: the technique and some applications of electropermeabilization. Biochem. J. 234, 497–506.
Scarpa, A. (1979) Transport across mitochondrial membranes, in Membrane Transport in Biology (Giebisch, G., Tosteson, D. C., and Ussing, H. H., eds.), Springer-Verlag, Berlin, pp. 263–355.
McCormack, J. G., and Denton, R. M. (1986) Ca2+ as a second messenger within mitochondria. TIBS 11, 258–262.
Andreason, G. L., and Evans, G. A. (1988) Introduction and expression of DNA molecules in eukaryotic cells by electroporation. BioTechniques 6, 650–660.
Nickoloff, J. A., and Reynolds, R. J. (1992) Electroporation-mediated gene transfer efficiency is reduced by linear plasmid carrier DNA. Anal. Biochem. 205, 237–243.
Potter, H. (1988) Electroporation in biology: methods, applications and instrumentation. Anal. Biochem. 174, 361–373.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Rols, M. P., and Teissie, J. (1990) Electropermeabilization of mammalian cells. Biophys. J. 58, 1089–1098.
Xie, T. D., Sun, L., and Tsong, T. Y. (1990) Study of mechanisms of electric field-induced DNA transfection. I. DNA entry by surface binding and diffusion through membrane pores. Biophys. J. 58, 13–19.
Knutson, J. C., and Yee, D. (1987) Electroporation: parameters affecting transfer of DNA into mammalian cells. Anal. Biochem. 164, 44–52.
Rols, M. P., Delteil, C., Serin, G., and Teissie, J. (1994) Temperature effects on electrotransfection of mammalian cells. Nucleic Acids Res. 22, 540.
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© 1995 Humana Press Inc.
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van den Hoff, M.J.B., Christoffels, V.M., Labruyère, W.T., Moorman, A.F.M., Lamers, W.H. (1995). Electrotransfection with “Intracellular” Buffer. In: Nickoloff, J.A. (eds) Animal Cell Electroporation and Electrofusion Protocols. Methods in Molecular Biology, vol 48. Humana Press. https://doi.org/10.1385/0-89603-304-X:185
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DOI: https://doi.org/10.1385/0-89603-304-X:185
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