Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

In vivo electrically mediated protein and gene transfer in murine melanoma

Nature Biotechnology volume 16, pages 168–171 (1998)Cite this article

Abstract

We show that efficient permeabilization of murine melanoma can be obtained in vivo by applying electric pulses. More than 80% of the cell population is affected as shown by the penetration of propidium iodide. A protein, (β-galactosidase, can be transferred and expressed into the cells by incorporating either the protein or a plasmid carrying the reporter gene with respective efficiencies of 20% and 4%. This is obtained by a direct injection of either the protein or the plasmid in the tumor, followed by the application of electric pulses with surface electrodes in contact with the skin. This approach is simple and safe to use, reproducible, and specific; moreover, it is potentially applicable to a wide variety of tissues, cell types, and animals.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Crystal, R.G. 1995. Transfer of genes to humans: early lessons and obstacles to success. Science 270:404-410.

    Article  CAS  Google Scholar 

  2. Culver, K.W. and Blaese, P.M. 1994. Gene therapy for cancer. Trends Genet. 10: 174–178.

    Article  Google Scholar 

  3. Herlyn, M. and Schuchter, L. 1996. Gene therapy–strategies and perspectives. The Skin Cancer Foundation Journal 14: 41.

    Google Scholar 

  4. Nabel, G.J., Nabel, E.G., Yang, Z.Y., Fox, B.A., Plautz, G.E., Gao, X., et al. 1993. Direct gene transfer with DNA–liposome complexes in melanoma: Expression, biologic activity and lack of toxicity in humans. Proc. Natl. Acad. Sci. USA 90: 11307–11311.

    Article  Google Scholar 

  5. Liu, Y., Mounkes, L.C., Liggit, H.D. Brown, C.S., Solodin, I., Heath, T.D., and Bebs, R.J. 1997. Factors influencing the efficiency of cationic liposome-mediated intravenous gene delivery. Bio/Technology 15: 167–173.

    Google Scholar 

  6. Neumann, E., Schaefer-Ridder, M., Wang, Y and Hofschneider, P.M. 1982. Gene transfer in mouse lyoma cells by electroporation in high electric fields. EMBO J. 1: 841–845.

    Article  Google Scholar 

  7. Wolf, H., Rols, M.P., Boldt, E. Neumann E., and Teissie, J. 1994. Control by pulse parameters of electric field mediated gene transfer in mammalian cells. Biophys. J., 66: 524–531.

    Article  Google Scholar 

  8. Tsong, T.Y. 1991. Electroporation of cell membranes. Biophys. J. 60: 297–306.

    Article  Google Scholar 

  9. Deuticke, B. and Schwister, K. 1989. Leaks induced by electrical breakdown in the erythrocyte membrane, pp. 127–148 in Electroporation and electrofusion in cell biology. Neumann, E., Sowers, A.E., and Jordan, C.A. (eds.), Plenum Press. New York.

    Chapter  Google Scholar 

  10. Marszalek, P., Liu, D.S ., and Tsong, T.Y. 1990. Schwan equation and transmem–brane potential induced by alternating electric field. Biophys. J. 58: 1053–1058.

    Article  Google Scholar 

  11. Genco, I., Gliozzi, A., Relini, A., Robello, M., and Scalas, E. 1993. Electroporation in symmetric and asymmetric membranes. Biochim. Biophys. Acta. 1149: 10–18.

    Article  Google Scholar 

  12. Teissie, J. and Rols,M.P. 1993. An experimental evaluation of the critical potential difference inducing cell membrane electropermeabilization. Biophys. J. 65: 409–413.

    Article  Google Scholar 

  13. Tekle, E., Astumian, R.D., and Chock, P.B. 1994. Selective and asymmetric molecular transport across electioporated membranes. Proc. Natl. Acad. Sci. USA 91: 11512–11516.

    Article  Google Scholar 

  14. Domenge, C., Orlowski, S., Luboinski, B., De Baere, T., Schwaab, G., Belehradek, J. Jr., et al. 1996. Antitumor Electrochemotherapy: new advances in the clinical protocol. Cancer 77: 956–962.

    Article  Google Scholar 

  15. Belehradek, M., Domenge, C.., Luboinski, B.., Orlowski, S., Belehradek, J. Jr ., and Mir, L.M. 1993. Electrochenfjotherapy, a new antitumor treatment: first clinical phase I–II trial. Cancer 72: 3612–3700.

    Article  Google Scholar 

  16. Mir, L.M., Orlowski, S..,, Belehradek, Jr., Teissie, J., Rols, M.P, Sersa, G., et al. 1995. Biomedical applications of electric pulses with special emphasis on antitumor electrochemotherapy. Bfoelectrochem. Bioenerg. 38: 203–207.

    Article  Google Scholar 

  17. Titomorov, A.V., Sukharev, S., and Kistanova, E. 1991. In vivo electroporation and stable transformation of skin cells of new born mice by plasmid DNA. Biochim. Biophys. Acta 1088: 131–134.

    Article  Google Scholar 

  18. Heller, R., Jaroszeski, M.,, Atkin, A., Moradpour, D., Gilbert, R., Wands, J., and Nicolau, C. 1996. In vivo gene electroinjection and expression in rat liver. FEBS Lett. 389: 225–228.

    Article  Google Scholar 

  19. Nishi, T, Yoshizato, K., Yamashiro, S., Takeshima, H., Sata, K., Hamada, K., et al. 1996. High–efficiency in vivo gene transfer using intraarterial plasmid DNA injection following in vivo electrqporation. Cancer Res. 56: 1050–1055.

    Google Scholar 

  20. Dagher, S.F., Conrad, S.E., Werner, E.A. and Patterson, R.J. Phenotypic conversion of TK deficient cells following electroporation of functional TK enzyme. Exp. Cell Res. 198, 36–42, 1991

    Article  Google Scholar 

  21. Goldman, C K., Soroceanu, L., Smith, N., Gillespie, Y., Shaw, W., Burgess, S., et al. 1997. In vitro and in vivo gene delivery mediated by a synthetic polycationic amino polymer. Bio/Technology 15: 462–466.

    Google Scholar 

  22. Rols, M.P. and Teissie, J. Flow cytometry quantification of electropermeabilization. In Methods in molecular biology: flow cytometry protocols. Jaroszeski, M.J. and Heller, R. (eds.), Humana Press Inc, Totowa, NJ. In press.

  23. Rols, M.P. and Teissié, J. 1990. Electropermeabilization of mammalian cells. Quantitative analysis of the phenomenon. Biophys.. 58: 1089–1098.

    Google Scholar 

  24. Rols, M.P., Femenia, P., and Teissié, J. 1995. Long lived macropinocytosis takes place in electropermeabilized mammalian cells. Biochem. Biophys. Res. Comm. 208: 26–35.

    Article  Google Scholar 

  25. Rols, M.P., Coulet, D., and Teissié, J. 1992. Highly efficient transfection of mammalian cells by electric field pulses. Application to large volumes of cell culture by using a flow system. Eur. J. Biochem. 206: 115–121.

    Article  Google Scholar 

  26. Rols, M.P., Delteil, C., Serin, G., and Teissie, J. 1994. Temperature effects on electrotransfection of mammalian cells. Nucl. Acids Res. 22: 540.

    Article  Google Scholar 

  27. Sersa, G., Cemazar, M., Semrov, D., and Miklavcic, D. 1996. Changing electrode orientation improves the efficacy of Electrochemotherapy of solid tumors in mice. Bioelectrochem. Bioenerg. 39: 61–66.

    Article  Google Scholar 

  28. Prautnitz,, M.R. 1996. The effects of electric current applied to skin: a review for transdermal drug delivery. Advanced Drug Delivery Reviews. 18: 395–425.

    Article  Google Scholar 

  29. Gilbert, R., Jaroszeski, M., and Heller, R. 1996. Novel electrodes designs for electrochemotherapy. Biochim. Biophys. Acta. 1334: 9–14.

    Article  Google Scholar 

  30. Heller, R., Jaroszeski, M., Leo-Massina, J., Perrot, R., Van Voorhis, N., Reintgen, D., and Gilbert, R. 1995. Treatment of B16 mouse melanoma with the combination of electropermeabilization and chemiotherapy. Bioelectrochem. Bioenerg. 36: 83–87.

    Article  Google Scholar 

  31. Bradford, M. 1976. A rapid and sensitive method for the quantitation of micro–gram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem. 72: 248–254.

    Article  Google Scholar 

Download references

Author information

Author notes

  1. Justin Teissie: e-mail: justin@lptf.biotoul.fr

Authors and Affiliations

  1. Institute of Pharmacology and Structural Biology, UPR 9062, 118 route de Narbonne, 31062, Toulouse cedex, France

    Marie-Pierre Rols, Christine Delteil, Muriel Golzio, Patrick Dumond, Suzie Cros & Justin Teissie

Authors
  1. Marie-Pierre Rols
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christine Delteil
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Muriel Golzio
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Patrick Dumond
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Suzie Cros
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Justin Teissie
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rols, MP., Delteil, C., Golzio, M. et al. In vivo electrically mediated protein and gene transfer in murine melanoma. Nat Biotechnol 16, 168–171 (1998). https://doi.org/10.1038/nbt0298-168

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0298-168

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing