Abstract
Aim:
To investigate the specific effects of Ca2+ on transgene expression during electroporation-mediated gene transfer in mice.
Methods:
Skeletal muscle and skin were subjected to in vivo electroporation with a luciferase reporter plasmid, with or without Ca2+ and various other ions.
Results:
For in vivo electroporation, the presence of just 10 mmol/L Ca2+ in the DNA solution drastically reduced the resulting transgene expression, to less than 5% of control values. Only Ca2+, not other ions, caused inhibition, and the effect was not tissue specific. More surprisingly, even when Ca2+ ions were delivered by electroporation before or after DNA administration, similar effects were still observed.
Conclusion:
The inhibitory effect of Ca2+ on in vivo gene transfer by electroporation is specific, ie, the inhibitory effect may be related to the cell membrane properties after electroporation and the subsequent resealing event.
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References
McMahon JM, Wells DJ . Electroporation for gene transfer to skeletal muscles: current status. BioDrugs 2004; 18: 155–65.
Li S, Benninger M . Applications of muscle electroporation gene therapy. Curr Gene Ther 2002; 2: 101–5.
Gardlik R, Palffy R, Hodosy J, Lukacs J, Turna J, Celec P . Vectors and delivery systems in gene therapy. Med Sci Monit 2005; 11: 110–21.
Gehl J, Mir LM . Determination of optimal parameters for in vivo gene transfer by electroporation, using a rapid in vivo test for cell permeabilization. Biochem Biophys Res Commun 1999; 261: 377–80.
Mir LM, Bureau MF, Gehl J, Rangara R, Rouy D, Caillaud JM, et al. High-efficiency gene transfer into skeletal muscle mediated by electric pulses. Proc Natl Acad Sci USA 1999; 96: 4262–7.
Lucas ML, Jaroszeski MJ, Gilbert R, Heller R . In vivo electroporation using an exponentially enhanced pulse: a new waveform. DNA Cell Biol 2001; 20: 183–8.
Lee MJ, Cho SS, Jang HS, Lim YS, You JR, Park J, et al. Optimal salt concentration of vehicle for plasmid DNA enhances gene transfer mediated by electroporation. Exp Mol Med 2002; 34: 265–72.
Suzuki T, Tsunekawa J, Murai A, Muramatsu T . Effect of CaCl2 concentration on the rate of foreign gene transfer and expression by in vivo electroporation in the mouse ovary. Int J Mol Med 2003; 12: 365–8.
Xu L, Zhao Y, Zhang Q, Li Y, Xu Y . Regulation of transgene expression in muscles by ultrasound-mediated hyperthermia. Gene Ther 2004; 11: 894–900.
Church PJ, Stanley EF . Single L-type calcium channel conductance with physiological levels of calcium in chick ciliary ganglion neurons. J Physiol 1996; 496: 59–68.
Neumann E, Kakorin S, Tsoneva I, Nikolova B, Tomov T . Calcium-mediated DNA adsorption to yeast cells and kinetics of cell transformation by electroporation. Biophys J 1996; 71: 868–77.
Gissel H, Clausen T . Ca2+ uptake and cellular integrity in rat EDL muscle exposed to electrostimulation, electroporation, or A23187. Am J Physiol Regul Integr Comp Physiol 2003; 285: 132–42.
Deng CX, Sieling F, Pan H, Cui J . Ultrasound-induced cell membrane porosity. Ultrasound Med Biol 2004; 30: 519–26.
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Project supported by a grant from the National 863 High Technology Research and Development Program of China (No 2002AA2Z3317) and a Shanghai Qimingxing grant to Yu-hong Xu (1404).
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Zhao, Yg., Lu, Hl., Peng, Jl. et al. Inhibitory effect of Ca2+ on in vivo gene transfer by electroporation. Acta Pharmacol Sin 27, 307–310 (2006). https://doi.org/10.1111/j.1745-7254.2006.00280.x
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DOI: https://doi.org/10.1111/j.1745-7254.2006.00280.x
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