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Nucleic Acids Research, 1993, Vol. 21, No. 15 3567-3573
© 1993

MOLECULAR BIOLOGY

Electroporation enhances c-myc antisense oligodeoxynucleotide efficacy

Raymond Bergan, Yvette Connell, Brigid Fahmy and Leonard Neckers

Clinical Pharmacology Branch, NCI, NIH Bethesda, MD 20892, USA

Received January 26, 1993. Revised May 14, 1993. Accepted May 14, 1993.

Obtaining high transfection efficiencies and achieving appropriate intracellular concentrations and localization are two of the most important barriers to the implementation of gene targeted therapy. The efficiency of endogenous uptake of oligodeoxynucleo-tides (ODNs) varies from cell type to cell type and may be a limiting factor of antisense efficacy. The use of electroporation to obtain high intracellular concentrations of a synthetic ODN in essentially 100% of viable cells is described. It is also shown that the transfected ODNs initially localize to the nucleus and remain there for at least 48 hours. The cellular trafficking of electroporated ODNs is shown to be an energy dependent process. Targeting of the c-myc proto-oncogene of U937 cells by electroporation of phosphorothioate-modified ODNs results in rapid and specific suppression of this gene at ODN concentrations much lower than would otherwise be required. This technique appears to be applicable to a variety of cell types and may represent a powerful new investigative tool as well as a promising approach to the ex vivo treatment of hematologic disorders.


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