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.
Highlights
Gene transfer depends on the effective delivery of nucleic acids (DNA, RNA) into cells or tissue and can be enabled by viral or nonviral delivery methods
Efficient silencing of enhanced green fluorescent protein (EGFP) after electrotransfer by high intensity pulsed electromagnetic field (HI-PEMF) was demonstrated by Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR)
EGFP mRNA levels were significantly reduced after HI-PEMF and conventional gene electrotransfer
Summary
Gene transfer depends on the effective delivery of nucleic acids (DNA, RNA) into cells or tissue and can be enabled by viral or nonviral delivery methods. One method for the nonviral delivery of genetic material (plasmid DNA, siRNA, and miRNA) into cells is electroporation. The use of electroporation as a gene delivery method (gene electrotransfer) was first reported in 1982 [3] and has since reached a broad spectrum of applications in different tissues, such as tumor, muscle, and skin [4,5,6,7]. Electroporation is used for the delivery of impermeant small molecules, such as dyes and drugs, into cells. Applying electroporation to facilitate intracellular delivery of cytotoxic drugs, such as bleomycin and cisplatin, Vaccines 2020, 8, 49; doi:10.3390/vaccines8010049 www.mdpi.com/journal/vaccines
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