Electric Field Orientation for Gene Delivery Using High-Voltage and Low-Voltage Pulses

Abstract

Electropermeabilization is a biological physical process in response to the presence of an applied electric field that is used for the transfer of hydrophilic molecules such as anticancer drugs or DNA across the plasma membranes of living cells. The molecular processes that support the transfer are poorly known. The aim of our study was to investigate the effect of high-voltage and low-voltage (HVLV) pulses in vitro with different orientations on cell permeabilization, viability and gene transfection. We monitored the permeabilization with unipolar and bipolar HVLV pulses with different train repetition pulses, showing that HVLV pulses increase cell permeabilization and cell viability. Gene transfer was also observed by measuring green fluorescent protein (GFP) expression. The expression was the same for HVLV pulses and electrogenotherapy pulses for in vitro experimentation. As the viability was better preserved for HVLV-pulsed cells, we managed to increase the number of GFP-expressing cells by up to 65% under this condition. The use of bipolar HVLV train pulses increased gene expression to a higher extent, probably by affecting a larger part of the cell surface.

Systematic optimization of square-wave electroporation conditions for bovine primary fibroblasts

“…Also, it is only at high electric field strengths that cell membrane progressively become less resistive [42]. The suitability of the high-voltage and low-voltage (HVLV) puls method [43] for bovine primary fibroblasts nessitates future research.…”

Efficient One-Step Knockout by Electroporation of Ribonucleoproteins Into Zona-Intact Bovine Embryos

“…While HV pulses are important to create pores for permeabilization, the LV pulses allow the DNA to be electrophoretically dragged into the cell ( Sukharev et al, 1992 ). In addition, bipolar LV pulses can increase the interaction between DNA and the membrane ( Faurie et al, 2004 ) and improve electrotransfer efficiency ( Orio et al, 2012 ). The combination of poring and transfer pulses could in part be responsible for the high rate of biallelic mutations observed in OCT4 gene (85%) compared to the Namula et al (2019) study which used only 3 poring pulses and obtained less than 5% biallelic mutations for the 15 V condition.…”

Irreversible electroporation enhances delivery of gemcitabine to pancreatic adenocarcinoma

“…Thus, the majority has used 6–8 high‐voltage pulses to deliver small molecules and perform gene transfer. Others, meanwhile, have favored a combination of high‐ and low‐voltage pulses to facilitate an electrophoretic effect on DNA . For treatment of superficial tumors, such as basal cell carcinoma and melanoma, voltage significantly impacts procedural discomfort and analgesia requirement, as pain from electroporation procedures has been shown to be proportional to absolute applied voltage .…”