Electroporation

Abstract

Electroporation is a simple, rapid and efficient technique for introducing DNA, RNA, proteins and other bioactive molecules into cells and tissues. Electroporation involves the creation of transient, electrically induced membrane pores through which molecules pass into cytoplasmic and nuclear compartments. Because electroporation involves a physical process acting on cell membranes, it is effective with a wide range of cell types including bacterial, fungal, plant and animal cells. Electroporation is used to transiently or stably increase gene expression through transfection of plasmidā€borne gene expression cassettes or decrease gene expression using antisense RNA, small interfering RNA or short hairpin RNA systems. Recent advances have led to new techniques for tissue and organ electroporation in living animals. These techniques are increasingly used in clinical applications including gene therapy, cancer drug delivery and vaccination.

Key Concepts:

  • Gene transfer is fundamental to molecular genetic analysis in bacteria, plant and animal cells.

  • Electroporation introduces a wide variety of bioactive molecules into cells, including DNA, RNA, siRNA, proteins, drugs and dyes.

  • Electroporation creates transient membrane pores through which molecules pass into cytoplasmic and nuclear compartments.

  • Electroporation is used to transiently or stably upregulate or downregulate gene expression.

  • Electroporation is used to make stable genetic changes to chromosomes including gene knockout and knockin.

  • Electroporation of molecules into tissues and organs of living organisms is driving novel clinical applications including gene therapy, drug delivery and vaccination.

  • The cytotoxic effects of electroporation are being used in tissue ablation including direct killing of tumours.

Keywords: DNA transfection; chromosomal modification; gene transfer; gene expression; gene targeting; gene therapy; protein transfection; drug delivery; cell fusion; cancer therapy

Figure 1.

Electroporation flowchart. Cells are harvested, rinsed to adjust ionic strength, DNA is added and the mixture is transferred to an electroporation cuvette, an electric pulse is applied, cells are transferred to culture dishes and antibiotic or other drug selection pressure is applied to select transfected cells.

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Further Reading

Lurquin PF (1997) Gene transfer by electroporation. Molecular Biotechnology 7: 5–35.

Nickoloff JA (1995) Animal Cell Electroporation and Electrofusion Protocols. Totowa, NJ: Humana Press.

Nickoloff JA (1995) Electroporation Protocols for Microorganisms. Totowa, NJ: Humana Press.

Nickoloff JA (1995) Plant Cell Electroporation and Electrofusion Protocols. Totowa, NJ: Humana Press.

Rubinsky B (2010) Irreversible Electroporation. New York, NY: Springer.

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How to Cite close
Nickoloff, Jac A(Oct 2010) Electroporation. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002654.pub2]