Electroporation of mammalian cells with a firefly luciferase expression plasmid: kinetics of transient expression differ markedly among cell types.
Electroporation in combination with the luciferase reporter system provides a rapid and sensitive means of determining transient expression from plasmid constructs in various mammalian cells. Conditions are described allowing efficient electroporation of several cell types with a commercial electroporation device. Measurements on extracts prepared at various times after electroporation showed a rapid rise in luciferase activity for up to 12 hr, which was followed, in certain cell types, by a rapid decline. With such cells, determinations made after 48-72 hr, as is conventional, would greatly underestimate expression of the luciferase reporter gene. Therefore, to observe optimum activity, the kinetics of expression must be determined for each cell type.
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Division of Medical Oncology, University of Colorado Health Sciences Center, Denver 80262.
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D002460:Cell Line; D006801:Humans; D066298:In Vitro Techniques; D008156:Luciferases / Q000235:genetics*; D010957:Plasmids; D014162:Transfection
D008156:Luciferases