Electric shock-mediated transfection of cells. Characterization and optimization of electrical parameters.
The effect of various parameters on the electric shock-mediated permeabilization and transfection of CHO cells has been investigated. Up to 70% of the cells can be maintained transiently permeable to erythrosin B for periods of at least 1 h at 20 degrees C. Electrical conditions optimal for transient permeabilization were also optimal for efficient DNA transfection by pSV2neo. However, the DNA must be present during exposure to the electric field for efficient transformation. The same requirement existed for voltage-induced DNA toxicity. The results suggest that DNA moves into the cells by electrophoresis, not by simple diffusion. Based on these observations a simple, rapid procedure for optimizing the conditions for electric shock-mediated DNA transfer into cells has been developed.
Winterbourne
D J
DJ
Department of Surgery, St George's Hospital Medical School, London, U.K.
Thomas
S
S
Hermon-Taylor
J
J
Hussain
I
I
Johnstone
A P
AP
D000818:Animals; D002021:Buffers; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival; D006224:Cricetinae; D004247:DNA / Q000032:analysis; D004558:Electric Stimulation; D004586:Electrophoresis; D009690:Nucleic Acid Conformation; D013696:Temperature; D013997:Time Factors; D014162:Transfection / Q000187:drug effects; D014357:Trypsin / Q000494:pharmacology
D002021:Buffers; D004247:DNA; D014357:Trypsin