High efficiency gene transfection by electroporation using a radio-frequency electric field.
In order to develop a safe and effective way to introduce exogenous genes into cells, we have experimented with a new method of electroporation which uses a radio-frequency (RF) electric field to permeabilize the cell membrane. This RF method has several advantages over the conventional electroporation method which uses a direct current (DC) field. We have shown that the RF electroporation method can be used to introduce marker genes into a wide variety of cell lines, including COS-M6, CV-1, CHO, 3T3 and hepatocytes, and is able to increase substantially the efficiency of gene transfection. (For example, the amount of DNA required for transfecting two million COS-M6 cells can be as low as 0.1 microgram). The transfection efficiency is shown to be affected by a number of factors, including cell type, field strength, pulse protocol and medium buffer. Because of its wide range of applications, high transfection efficiency and lack of harmful side-effect, the RF electroporation method would be particularly useful for introducing genes into human cells for gene therapy.
Chang
D C
DC
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX 77030.
Gao
P Q
PQ
Maxwell
B L
BL
D000818:Animals; D002021:Buffers; D002452:Cell Count; D002460:Cell Line; D002463:Cell Membrane Permeability; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004247:DNA / Q000235:genetics; D004560:Electricity; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005819:Genetic Markers; D009994:Osmolar Concentration; D011846:Radio Waves; D013997:Time Factors; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D002021:Buffers; D005819:Genetic Markers; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase