Comparison between direct and reverse electroporation of cells in situ: a simulation study

Abstract

The discovery of the human genome has unveiled new fields of genomics, transcriptomics, and proteomics, which has produced paradigm shifts on how to study disease mechanisms, wherein a current central focus is the understanding of how gene signatures and gene networks interact within cells. These gene function studies require manipulating genes either through activation or inhibition, which can be achieved by temporarily permeabilizing the cell membrane through transfection to deliver cDNA or RNAi. An efficient transfection technique is electroporation, which applies an optimized electric pulse to permeabilize the cells of interest. When the molecules are applied on top of seeded cells, it is called “direct” transfection and when the nucleic acids are printed on the substrate and the cells are seeded on top of them, it is termed “reverse” transfection. Direct transfection has been successfully applied in previous studies, whereas reverse transfection has recently gained more attention in the context of high‐throughput experiments. Despite the emerging importance, studies comparing the efficiency of the two methods are lacking. In this study, a model for electroporation of cells in situ is developed to address this deficiency. The results indicate that reverse transfection is less efficient than direct transfection. However, the model also predicts that by increasing the concentration of deliverable molecules by a factor of 2 or increasing the applied voltage by 20%, reverse transfection can be approximately as efficient as direct transfection.

Translocation of silica nanospheres through giant unilamellar vesicles (GUVs) induced by a high frequency electromagnetic field

“…52 The collapse of thus-formed bubbles may also lead to localised sharp increase in temperature, 53 with the latter having capacity to catalyse chemical reactions, such as hydrolysis of molecules comprising lipid bilayers. 54 The permeation process described in this study differs from electroporation which is another technique used in breaching the cell membrane barrier 55,56 which results in increasing cell membrane permeability. 57 Electroporation can be applied to release nucleic acids, intracellular proteins and other metabolites out of the cell for analysis purposes.…”

“…55 The mechanism behind electroporation is the increase in transmembrane potential (TMP) of a cell above 0.2-1 V, 55 irrespective of the cell type leading to reversible pore formation. These electro-pores allow DNA or large molecules to enter the cell [55][56][57][58][59][60] by the application of external pulsed electric elds (PEFs). 61 The method uses pulsed electric eld to cause permeability of cell membranes.…”

“…58 Therapies based on electroporation involved placing electrodes around or within a target tissue while delivering a series of 8-100 short ($100 ms) electric pulses of high voltage ($1000-3000 V); 58 the membrane can return to its intact state aer pulse withdrawal. 55,56,62 Cell membrane permeabilization resulting from the exposure to the EMF of 18 GHz can be classied as a different phenomenon compared to cell poration phenomena achieved using other techniques, including mechanical stress, sonoporation, electroporation and photoporation. 57,60,63 An EMF-induced change in the hydration of H 3 O + and OH À typically present in water, specically the removal of the hydration shell, may increase their chemical activity, 64,65 leading to solution structuring, e.g., cluster formation, and reactive species production, 66 as demonstrated by retained activity of the solution aer EMF is removed.…”

Investigation of ac-magnetic field stimulated nanoelectroporation of magneto-electric nano-drug-carrier inside CNS cells

“…Considering the required mesh density for each domain in COMSOL, inclusion of an extra domain with 5 nm small region for cell membrane thickness is not only impractical but also computationally expensive. Hence, cell membrane was modelled as boundary condition in the simulation3839. The “contact impedance” boundary condition, available in COMSOL under the electric currents of AC/DC module, is meant to approximate a thin layer of material that impedes the flow of current normal to the boundary, but does not introduce any additional conduction path tangential to the boundary.…”

Localization of nanospheres in pheochromocytoma-like cells following exposure to high-frequency electromagnetic fields at 18 GHz

“…We previously showed that PC 12 cells remained viable after transient membrane permeability [13]. In this work, we studied the viability of PC 12 cells after 2 and 4 days post-EMF exposure (the cell viability and morphology after day 1 and 5 are shown in electronic supplementary material, figure royalsocietypublishing.org/journal/rsos R. Soc.…”