Improvement in Electrotransfection of Cells Using Carbon-Based Electrodes
- 189 Downloads
Electrotransfection has been widely used as a versatile, non-viral method for gene delivery. However, electrotransfection efficiency (eTE) is still low and unstable, compared to viral methods. To understand potential mechanisms of the problems, we investigated effects of electrode materials on eTE and viability of mammalian cells. Data from the study showed that commonly used metal electrodes generated a significant amount of particles during application of pulsed electric field, which could cause precipitation of plasmid DNA from solutions, thereby reducing eTE. For aluminum electrodes, the particles were composed of aluminum hydroxide and/or aluminum oxide, and their median sizes were 300–400 nm after the buffer was pulsed 4–8 times at 400 V cm−1, 5 ms duration and 1 Hz frequency. The precipitation could be prevented by using carbon (graphite) electrodes in electrotransfection experiments. The use of carbon electrodes also increased cell viability. Taken together, the study suggested that electrodes made of electrochemically inert materials were desirable for electrotransfection of cells in vitro.
KeywordsElectrotransfection Electro-gene delivery Electroporation Carbon electrodes DNA precipitation
The work was supported partly by grants from National Institutes of Health (GM098520) and National Science Foundation (BES-0828630).
Conflict of Interest
Chun-Chi Chang, Mao Mao, Yang Liu, Mina Wu, Tuan Vo-Dinh, and Fan Yuan declare that they have no conflict of interest.
No human or animal studies were carried out by the authors for this article.
- 8.Heller, R., and L. C. Heller. Gene electrotransfer clinical trials. Adv. Genet. 89:235–262, 2015.Google Scholar
- 13.Kooijmans, S. A. A., S. Stremersch, K. Braeckmans, S. C. De Smedt, A. Hendrix, M. J. A. Wood, R. M. Schiffelers, K. Raemdonck, and P. Vader. Electroporation-induced siRNA precipitation obscures the efficiency of siRNA loading into extracellular vesicles. J. Control. Release 172:229–238, 2013.CrossRefGoogle Scholar
- 15.Loomis-Husselbee, J. W., P. J. Cullen, R. F. Irvine, and A. P. Dawson. Electroporation can cause artefacts due to solubilization of cations from the electrode plates. Aluminum ions enhance conversion of inositol 1,3,4,5-tetrakisphosphate into inositol 1,4,5-trisphosphate in electroporated L1210 cells. Biochem. J. 277(3):883–885, 1991.CrossRefGoogle Scholar
- 18.Neumann, E., M. Schaefer-Ridder, Y. Wang, and P. H. Hofschneider. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1:841–845, 1982.Google Scholar
- 19.Nishi, T., K. Yoshizato, S. Yamashiro, H. Takeshima, K. Sato, K. Hamada, I. Kitamura, T. Yoshimura, H. Saya, J. Kuratsu, and Y. Ushio. High-efficiency in vivo gene transfer using intraarterial plasmid DNA injection following in vivo electroporation. Cancer Res. 56:1050–1055, 1996.Google Scholar
- 26.Touchard, E., M. Berdugo, P. Bigey, M. El Sanharawi, M. Savoldelli, M.-C. Naud, J.-C. Jeanny, and F. Behar-Cohen. Suprachoroidal electrotransfer: a nonviral gene delivery method to transfect the choroid and the retina without detaching the retina. Mol. Therapy 20:1559–1570, 2012.CrossRefGoogle Scholar
- 28.Wolff, J. A., and V. Budker. The mechanism of naked DNA uptake and expression. Adv. Genet. 54:1–20, 2015.Google Scholar
- 30.Xu, Z. P., T. L. Walker, K.-L. Liu, H. M. Cooper, G. Q. M. Lu, and P. F. Bartlett. Layered double hydroxide nanoparticles as cellular delivery vectors of supercoiled plasmid DNA. Int. J. Nanomed. 2:163–174, 2007.Google Scholar