Dependence of DNA electrotransfer into cells in vitro on cell electroporation and DNA electrophoresis

Abstract

Electroporation mostly refers to the technique of permeabilizing cell membranes by applying short and intense electric pulses across a cell, such that the barrier function of the membrane is temporally disturbed. This allows various molecules, including plasmid DNA to pass across the membrane and induce their biological effects. The mechanism of the translocation of plasmid DNA through the permeabilized membrane is different from simple diffusion and remains poorly understood. In this in vitro study we used two types of pulses, high voltage, short duration (HV) and low voltage short duration (LV) pulses to evaluate cell electrotransfection efficiency in dependence of cell permeabilization (HV pulse) and DNA electrophoresis (LV pulse). Our results showed that DNA electrotransfer was mainly dependent on perneabilizing HV pulse. Differently from in vivo studies, this study showed that contribution of LV pulse to DNA electrotransfer is negligible.