Electrotransfer of siRNA to Silence Enhanced Green Fluorescent Protein in Tumor Mediated by a High Intensity Pulsed Electromagnetic Field

Abstract

The contactless high intensity pulsed electromagnetic field (HI-PEMF)-induced increase of cell membrane permeability is similar to conventional electroporation, with the important difference of inducing an electric field non-invasively by exposing a treated tissue to a time-varying magnetic field. Due to the limited number of studies in the field of electroporation induced by HI-PEMF, we designed experiments to explore the feasibility of such a contactless delivery technique for the gene electrotransfer of nucleic acids in tissues in vivo. By using HI-PEMF for gene electrotransfer, we silenced enhanced green fluorescent protein (EGFP) with siRNA molecules against EGFP in B16F10-EGFP tumors. Six days after the transfer, the fluorescent tumor area decreased by up to 39% as determined by fluorescence imaging in vivo. In addition, the silencing of EGFP to the same extent was confirmed at the mRNA and protein level. The results obtained in the in vivo mouse model demonstrate the potential use of HI-PEMF-induced cell permeabilization for gene therapy and DNA vaccination. Further studies are thus warranted to improve the equipment, optimize the protocols for gene transfer and the HI-PEMF parameters, and demonstrate the effects of HI-PEMF on a broader range of different normal and tumor tissues.

Determination of the Impact of High-Intensity Pulsed Electromagnetic Fields on the Release of Damage-Associated Molecular Pattern Molecules

“…In 2012, a magnetic nerve stimulator was applied for the permeabilization of cells in guinea pig skin in vivo to enhance uptake and expression of GFP plasmid DNA [8]. We later demonstrated that HI-PEMF can be applied for delivering siRNA molecules to silence enhanced green fluorescent protein (EGFP) in B16F10-EGFP mouse tumors in vivo [40]. Since siRNA delivery is a promising gene therapy approach for inactivating oncogenes and tumor suppressor genes involved in cancer disease [41], the results obtained demonstrate the potential use of HI-PEMF for cancer therapy.…”

A Modified Marx Generator Circuit with Enhanced Tradeoff between Voltage and Pulse Width for Electroporation Applications

“…Electroporation has been useful in various fields such as biotechnology, medicine, genetics, drug kinetics, and dynamics [3,[5][6][7][8]. The electroporator creates pores if the pulse voltage is within a precise range [9] and faces a challenge in the application of a high voltage for a short duration of time (milliseconds to nanoseconds).The high pulse period causes cell death; hence, it is dangerous.…”

High-Intensity Pulsed Electromagnetic Field-Mediated Gene Electrotransfection In Vitro

“…The introduction of small biomolecules, nucleic acids, proteins, synthetic nanomaterials and drugs into cells is a powerful means of monitoring and deciphering cellular behavior and influencing cellular fate and a cell’s biological functions [ 1 , 2 , 3 ]. The effects of high-intensity pulsed electromagnetic fields (HI-PEMF) on the permeability of cell membranes have already been observed: in vitro in mammalian cells and microorganisms [ 4 , 5 ] and in vivo in small animals [ 6 , 7 , 8 , 9 ]. Various molecules have been used to demonstrate cell membrane permeabilization by HI-PEMF, including propidium iodide [ 10 , 11 ], YO-PRO-1 [ 10 ], lucifer yellow [ 12 ], cisplatin, bleomycin [ 6 ] and calcein [ 13 ].…”