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Electroporation is a physical method to induce the uptake of therapeuticdrugs and DNA, by eukaryotic cells and tissues. The phenomena behindelectro-mediated membrane permeabilization to plasmid DNA have been shown to besignificantly more complex than those for small molecules. Small moleculescross the permeabilized membrane by diffusion whereas plasmid DNA firstinteracts with the electropermeabilized part of the cell surface, forminglocalized aggregates. The dynamics of this process is still poorly understoodbecause direct observations have been limited to scales of the order ofseconds. Here, cells are electropermeabilized in the presence of plasmid DNAand monitored with a temporal resolution of 2 ms. This allows us to show thatduring the first pulse application, plasmid complexes, or aggregates, start toform at distinct sites on the cell membrane. FRAP measurements show that thepositions of these sites are remarkably immobile during the application offurther pluses. A theoretical model is proposed to explain the appearance ofdistinct interaction sites, the quantitative increase in DNA and also theirimmobility leading to a tentative explanation for the success ofelectro-mediated gene delivery.

Electromediated formation of DNA complexes with cell membranes and its consequences for gene delivery