Improved intracellular delivery of oligonucleotides by square wave electroporation.
Prior studies have shown that electroporation is a simple and effective method for the introduction of oligonucleotides (ODN) into cells. In ex vivo bone marrow purging models, electroporation of ODN into cells has been associated with selective killing of human neoplastic cells while sparing hematopoietic stem cells. Prior studies used conventional electroporation methods (i.e., exponential decay) to introduce ODN into cells. Square wave electroporation allows the delivery of a more defined and regulated electrical pulse and is associated with high transfection efficiencies in a variety of systems. The current study was undertaken to determine whether square wave electroporation was more effective than exponential decay electroporation for the delivery of ODN into hematopoietic cells. Using fluorescein-tagged ODN and K562, chronic myelogenous leukemia (CML) cells, higher transfection rates were observed after square wave electroporation. In addition, c-myc antisense ODN were more effective in reducing c-myc protein when introduced by square wave electroporation, as compared with introduction by exponential decay electroporation. Square wave electroporation is thus identified as the optimal method for delivering ODN into hematopoietic cells.
Department of Medicine, Northwestern University Medical School, and the Lurie Cancer Center of Northwestern University, Chicago, IL 60611-3008, USA.
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Liu
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Bergan
D001483:Base Sequence; D001854:Bone Marrow Cells / Q000378:metabolism; D002470:Cell Survival; D018274:Electroporation / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D016259:Genes, myc; D006801:Humans; D020014:K562 Cells; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D016271:Proto-Oncogene Proteins c-myc / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D005456:Fluorescent Dyes; D020319:Oligodeoxyribonucleotides, Antisense; D016271:Proto-Oncogene Proteins c-myc; D016650:Fluorescein-5-isothiocyanate