Global InformationElectroporation information
Electroporation, or electropermeabilization, is a technique in which an electrical field is applied to cells in order to increase the permeability of the cell membrane. This may allow chemicals, drugs, electrode arrays or DNA to be introduced into the cell (also called electrotransfer).[1][2][3][4]
In microbiology, the process of electroporation is often used to transform bacteria, yeast, or plant protoplasts by introducing new coding DNA. If bacteria and plasmids are mixed together, the plasmids can be transferred into the bacteria after electroporation, though depending on what is being transferred, cell-penetrating peptides or cell squeeze could also be used. Electroporation works by passing thousands of volts (~8 kV/cm) across suspended cells in an electroporation cuvette.[2] Afterwards, the cells have to be handled carefully until they have had a chance to divide, producing new cells that contain reproduced plasmids. This process is approximately ten times more effective in increasing cell membrane's permeability than chemical transformation, although many laboratories lack the specialized equipment needed for electroporation.[5][6][verification needed]
Electroporation is also highly efficient for the introduction of foreign genes into tissue culture cells, especially mammalian cells. For example, it is used in the process of producing knockout mice, as well as in tumor treatment, gene therapy, and cell-based therapy. The process of introducing foreign DNA into eukaryotic cells is known as transfection. Electroporation is highly effective for transfecting cells in suspension using electroporation cuvettes. Electroporation has proven efficient for use on tissues in vivo, for in utero applications as well as in ovo transfection. Adherent cells can also be transfected using electroporation, providing researchers with an alternative to trypsinizing their cells prior to transfection. One downside to electroporation, however, is that after the process the gene expression of over 7,000 genes can be affected.[7] This can cause problems in studies where gene expression has to be controlled to ensure accurate and precise results.
Although bulk electroporation has many benefits over physical delivery methods such as microinjections and gene guns, it still has limitations, including low cell viability. Miniaturization of electroporation has been studied, leading to microelectroporation and nanotransfection of tissue utilizing electroporation-based techniques via nanochannels to minimally invasively deliver cargo to the cells.[8] [9]
Electroporation has also been used as a mechanism to trigger cell fusion. Artificially induced cell fusion can be used to investigate and treat different diseases, like diabetes,[10][11][12] regenerate axons of the central nerve system,[13] and produce cells with desired properties, such as in cell vaccines for cancer immunotherapy.[14] However, the first and most known application of cell fusion is production of monoclonal antibodies in hybridoma technology, where hybrid cell lines (hybridomas) are formed by fusing specific antibody-producing B lymphocytes with a myeloma (B lymphocyte cancer) cell line.[15]
- ^ Muralidharan A, Boukany PE (December 2023). "Electrotransfer for nucleic acid and protein delivery". Trends in Biotechnology. doi:10.1016/j.tibtech.2023.11.009. PMID 38102019.
- ^ a b Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH (1982). "Gene transfer into mouse lyoma cells by electroporation in high electric fields". The EMBO Journal. 1 (7): 841–845. doi:10.1002/j.1460-2075.1982.tb01257.x. PMC 553119. PMID 6329708.
- ^ Jimbo Y, Sasaki D, Ohya T, Lee S, Lee W, Arab Hassani F, et al. (September 2021). "An organic transistor matrix for multipoint intracellular action potential recording". Proceedings of the National Academy of Sciences of the United States of America. 118 (39): e2022300118. Bibcode:2021PNAS..11822300J. doi:10.1073/pnas.2022300118. PMC 8488610. PMID 34544852. S2CID 237584521.
- ^ Chang DC (2006-09-15). "Electroporation and Electrofusion". In Meyers RA (ed.). Encyclopedia of Molecular Cell Biology and Molecular Medicine. Wiley-VCH Verlag GmbH & Co. KGaA. doi:10.1002/3527600906.mcb.200300026. ISBN 9783527600908.
- ^ Liu J, Chang W, Pan L, Liu X, Su L, Zhang W, et al. (December 2018). "An Improved Method of Preparing High Efficiency Transformation Escherichia coli with Both Plasmids and Larger DNA Fragments". Indian Journal of Microbiology. 58 (4): 448–456. doi:10.1007/s12088-018-0743-z. PMC 6141401. PMID 30262955.
- ^ Sugar IP, Neumann E (May 1984). "Stochastic model for electric field-induced membrane pores. Electroporation". Biophysical Chemistry. 19 (3): 211–225. doi:10.1016/0301-4622(84)87003-9. PMID 6722274.
- ^ Anne Trafton (2 February 2016). "Cell squeezing enhances protein imaging". MIT News Office.
- ^ Gallego-Perez D, Pal D, Ghatak S, Malkoc V, Higuita-Castro N, Gnyawali S, et al. (October 2017). "Topical tissue nano-transfection mediates non-viral stroma reprogramming and rescue". Nature Nanotechnology. 12 (10): 974–979. Bibcode:2017NatNa..12..974G. doi:10.1038/nnano.2017.134. PMC 5814120. PMID 28785092.
- ^ Muralidharan A, Pesch GR, Hubbe H, Rems L, Nouri-Goushki M, Boukany PE (October 2022). "Microtrap array on a chip for localized electroporation and electro-gene transfection". Bioelectrochemistry. 147: 108197. doi:10.1016/j.bioelechem.2022.108197. PMID 35810498.
- ^ McClenaghan NH (May 2007). "Physiological regulation of the pancreatic {beta}-cell: functional insights for understanding and therapy of diabetes". Experimental Physiology. 92 (3): 481–96. doi:10.1113/expphysiol.2006.034835. PMID 17272356. S2CID 22548866.
- ^ Yanai G, Hayashi T, Zhi Q, Yang KC, Shirouzu Y, Shimabukuro T, et al. (2013). "Electrofusion of mesenchymal stem cells and islet cells for diabetes therapy: a rat model". PLOS ONE. 8 (5): e64499. Bibcode:2013PLoSO...864499Y. doi:10.1371/journal.pone.0064499. PMC 3665804. PMID 23724055.
- ^ McCluskey JT, Hamid M, Guo-Parke H, McClenaghan NH, Gomis R, Flatt PR (June 2011). "Development and functional characterization of insulin-releasing human pancreatic beta cell lines produced by electrofusion". The Journal of Biological Chemistry. 286 (25): 21982–92. doi:10.1074/jbc.M111.226795. PMC 3121343. PMID 21515691.
- ^ Sretavan DW, Chang W, Hawkes E, Keller C, Kliot M (October 2005). "Microscale surgery on single axons". Neurosurgery. 57 (4): 635–46, discussion 635–46. doi:10.1227/01.NEU.0000175545.57795.ac. PMID 16239875. S2CID 196411777.
- ^ Takakura K, Kajihara M, Ito Z, Ohkusa T, Gong J, Koido S (March 2015). "Dendritic-tumor fusion cells in cancer immunotherapy". Discovery Medicine. 19 (104): 169–74. PMID 25828520.
- ^ Trontelj K, Rebersek M, Kanduser M, Serbec VC, Sprohar M, Miklavcic D (November 2008). "Optimization of bulk cell electrofusion in vitro for production of human-mouse heterohybridoma cells". Bioelectrochemistry. 74 (1): 124–9. doi:10.1016/j.bioelechem.2008.06.003. PMID 18667367.