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Nucleic Acids Research, 1989, Vol. 17, No. 10 3959-3971
© 1989

CHEMISTRY

Enhanced transfection efficiency and improved cell survival after electroporation of G2/M-synchronized cells and treatment with sodium butyrate

Samuel Goldstein*,+, C.Michael Fordis and Bruce H. Howard

Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health Bethesda, MD 20892, USA

*To whom correspondence should be addressed

Received January 10, 1989. Revised April 12, 1989. Accepted April 12, 1989.

To achieve high transfection efficiency in human fibroblasts with good preservation of proiiferative capacity we developed an electroporation procedure that combines two distinct modalities: use of recipient cells synchronized in the late G2/mitotic phase of the cell cycle and treatment of cells post-electroporation with 5 mM butyrate. This combination enabled reduction of plasmid DNA concentration and electroporation voltage, both associated with cytotoxicity, while greatly enhancing transfection efficiencies. Although the method was primarily developed for transient expression it was also found to improve stable expression. This procedure should have wide applicability, particularly in studies seeking to identify DNA sequences that lead to inhibition of DNA synthesis and proliferation in human fibroblasts and other cells refractory to transfection.

+Present address: Departments of Medicine, Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences and Geriatnc Research Education and Clinical Center, John L.McClellan Memorial Veterans Hospital, 151 Research, Little Rock, AR 72205, USA


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