Gene transfer into mouse lyoma cells by electroporation in high electric fields.
Electric impulses (8 kV/cm, 5 microseconds) were found to increase 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 selection medium. At 20 degrees C after the application of three successive electric impulses followed by 10 min to allow DNA entry there result 95 (+/- 3) transformants per 10(6) cells and per 1.2 micrograms 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 electric 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.
D000818:Animals; D003001:Cloning, Molecular; D004558:Electric Stimulation; D005796:Genes; D005814:Genes, Viral; D007700:Kinetics; D007739:L Cells / Q000502:physiology; D051379:Mice; D010957:Plasmids; D018139:Simplexvirus / Q000235:genetics; D013816:Thermodynamics; D013937:Thymidine Kinase / Q000235:genetics
D013937:Thymidine Kinase