Nanosecond Electric Pulse Effects on Gene Expression

The Journal of Membrane Biology - Tập 246 - Trang 851-859 - 2013
Louise Chopinet1,2, Tina Batista-Napotnik3, Audrey Montigny1,2, Matej Rebersek3, Justin Teissié1,2, Marie-Pierre Rols1,2, Damijan Miklavčič3
1CNRS, IPBS - UMR 5089, Toulouse Cedex 4, France
2UT1, UTM, LAAS, ITAV, UPS, INSA, INP, ISAE, Université de Toulouse, Toulouse Cedex 4, France
3Faculty of Electrical Engineering, University of Ljubljana, Ljubljana, Slovenia

Tóm tắt

Gene electrotransfection using micro- or millisecond electric pulses is a well-established method for safe gene transfer. For efficient transfection, plasmid DNA has to reach the nucleus. Shorter, high-intensity nanosecond electric pulses (nsEPs) affect internal cell membranes and may contribute to an increased uptake of plasmid by the nucleus. In our study, nsEPs were applied to Chinese hamster ovary (CHO) cells after classical gene electrotransfer, using micro- or millisecond pulses with a plasmid coding the green fluorescent protein (GFP). Time gaps between classical gene electrotransfer and nsEPs were varied (0.5, 2, 6 and 24 h) and three different nsEP parameters were used: 18 ns-10 kV/cm, 10 ns-40 kV/cm and 15 ns-60 kV/cm. Results analyzed by either fluorescence microscopy or flow cytometry showed that neither the percentage of electrotransfected cells nor the amount of GFP expressed was increased by nsEP. All nsEP parameters also had no effects on GFP fluorescence intensity of human colorectal tumor cells (HCT-116) with constitutive expression of GFP. We thus conclude that nsEPs have no major contribution to gene electrotransfer in CHO cells and no effect on constitutive GFP expression in HCT-116 cells.

Tài liệu tham khảo

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