Strategy for increased efficiency of transfection in human cell lines using radio frequency electroporation.
Traditional electroporation devices use direct current electric fields to stimulate the uptake of oligonucleotides, plasmids, short peptides, and proteins into a variety of cell types. A variation of this widely used technique is now available which relies on radio frequency (RF) electrical pulses. This oscillating type of electrical field reportedly elicits greater uptake of plasmid DNA across the plasma membrane. We evaluated a protocol for RF electroporation of the a human embryonic kidney cell line and a Burkitt's lymphoma (BL) cell line for effeciency of transfection by RF electroporation. The plasmid EGFP, which codes for the widely used fusion protein, enhanced green fluorescent protein (EGFP), was used as a reporter of plasmid uptake after transfections. Transfection efficiency consistently increased approximately 30% from that typically obtained with conventional DC type electroporation and was accompanied by greater survivability of cells. Additionally, in some instances, percent transfection efficiency increased to over 70%. Thus, RF electroporation represents an improved methodology for transfection of human cell lines. Moreover, the RF protocol is simple to incorporate in laboratories already utilizing conventional electroporation devices and techniques.
Zald
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PB
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor 48109-0934, USA.
Cotter
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MA
Robertso
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ES
D002051:Burkitt Lymphoma; D002460:Cell Line; D002470:Cell Survival; D018274:Electroporation; D004622:Embryo, Mammalian; D006801:Humans; D007668:Kidney; D008856:Microscopy, Fluorescence; D011846:Radio Waves; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured