The Role of Electrophoresis in Gene Electrotransfer |
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Authors: | M Pavlin K Flisar M Kandu?er |
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Institution: | (1) Faculty of Electrical Engineering, University of Ljubljana, Tržaška 25, Ljubljana, Slovenia |
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Abstract: | Gene electrotransfer is an established method for gene delivery which uses high-voltage pulses to increase the permeability
of a cell membrane and enables transfer of genes. Poor plasmid mobility in tissues is one of the major barriers for the successful
use of gene electrotransfer in gene therapy. Therefore, we analyzed the effect of electrophoresis on increasing gene electrotransfer
efficiency using different combinations of high-voltage (HV) and low-voltage (LV) pulses in vitro on CHO cells. We designed
a special prototype of electroporator, which enabled us to use only HV pulses or combinations of LV + HV and HV + LV pulses.
We used optimal plasmid concentrations used in in vitro conditions as well as lower suboptimal concentrations in order to
mimic in vivo conditions. Only for the lowest plasmid concentration did the electrophoretic force of the LV pulse added to
the HV pulse increase the transfection efficiency compared to using only HV. The effect of the LV pulse was more pronounced
for HV + LV, while for the reversed sequence, LV + HV, there was only a minor effect of the LV pulse. For the highest plasmid
concentrations no added effect of LV pulses were observed. Our results suggest that there are different contributing effects
of LV pulses: electrophoretically increased contact of DNA with the membrane and increased insertion of DNA into permeabilized
cell membrane and/or translocation due to electrophoretic force, which appears to be the dominant effect. |
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