Effect of time duration between HV and LV pulses on DNA electrotransfection efficiency in vitro

Abstract

Gene electrotransfer is a promising and expanding technique for various clinical applications, such as DNA vaccination and gene therapy, which is becoming a suitable alternative to virus-based methods. The main focus of our study in this field was to improve gene electrotransfection efficiency by combining high voltage (HV) and low-voltage (LV) electric pulses [1,2]. We investigated the effect of time duration between high-voltage (HV) and low-voltage (LV) electric pulses on plasmid DNA electrotransfer efficiency to CHO cells in vitro. Green fluorescent protein (GFP) coding plasmid was used in the experiments. We evaluated electrotransfection efficiency for GFP coding plasmid using flow cytometry to determine the percentage of GFP positive cells and the fluorescence intensity in them. The best electrotransfection efficiency was obtained with minimum time duration between HV and LV pulses. However, cell viability at lowest time duration between HV and LV pulses was also significantly decreased.