Gene transfer into mouse lyoma cells by electroporation in high electric fields
Neumann, E.; Schaefer-Ridder, M.; Wang, Y.; Hofschneider, P.H.
EMBO Journal 1(7): 841-845
1982
ISSN/ISBN: 0261-4189 PMID: 6329708 Document Number: 434
Electric impulses (8 kV/cm, 5 .mu.S) increased greatly the uptake of DNA into cells. When linear or circular plasmid DNA containing the herpes simplex thymidine kinase (TK) gene is added to a suspension of mouse L cells deficient in the TK gene and the cells are then exposed to electric fields, stable transformants are formed that survive in the HAT [hypoxanthine-aminopterin-thymidine] selection medium. At 20.degree. C after the application of 3 successive electric impulses followed by 10 min to allow DNA entry there resulted 95 (.+-. 3) transformants per 106 cells and per 1.2 .mu.g DNA. Compared with biochemical techniques, the electric field method of gene transfer is very simple, easily applicable, and very efficient. Because the mechanism of DNA transport through cell membranes is not known, a simple physical model for the enhanced DNA penetration into cells in high electic fields is proposed. According to this electroporation model the interaction of the external electric field with the lipid dipoles of a pore configuration induces and stabilizes the permeation sites and thus enhances cross membrane transport.
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