Gene therapy is an advanced treatment approach in medicine. The process of introducing foreign genetic material into target cells is called transfection. Among non-viral methods, electroporation is a physical method in which the permeability of the cell membrane is increased by means of an applied external electric field, thus enabling delivery of molecules into the cells. Many chemical methods, such as cationic lipids are also used as in vitro carriers of nucleic acids. For transfection to be successful many different parameters need to be considered. In addition to the transfection efficiency and the expression level of reporter protein, cell survival and activation of the host immune system are also important. The aim of our study was to determine the transfection efficiency after gene electrotransfer by using electroporation and K2 transfection system as a transfection method mediated by cationic lipids. We have shown that the K2 transfection system represents a better choice for in vitro delivery of plasmid DNA into murine melanoma cells than electroporation. The number of transfected cells after transfection with K2 transfection system was statistically significantly higher for low metastatic potential B16F1 melanoma, while the fluorescence intensity was statistically significantly higher for high metastatic potential B16F10 melanoma compared to gene electrotransfer. In addition, K2 transfection system was completely nontoxic and did not activate the cytosolic sensor DDX60, which can lead to activation of signalling pathways and induce apoptosis. After gene electrotransfer we observed an increased expression of a cytosolic sensor DDX60, a decrease in cell survival, and differences in transfection efficiency depended on the cell line. K2 transfection system was also more successful than the control method lipofection.
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