Efficient gene transfer by sequential treatment of mammalian cells with DEAE-dextran and deoxyribonucleic acid.
A variation of the classical DEAE-dextran method of gene transfer was developed for efficient transfection of HeLa cells with plasmid DNA. A brief exposure of the cells to medium containing DEAE-dextran was found to be sufficient for subsequent uptake of pRSVcat and to be superior to cocultivation of the cells with DEAE-dextran plus DNA. This sequential method of gene transfer is nontoxic and yielded up to 60% of HeLa cells positive for a surface protein encoded by the transfected sequence. The implications of this sequential transfection technique regarding the mechanisms of gene transfer are discussed.
Holter
W
W
Laboratory of Molecular Biology, National Cancer Institute, Bethesda, Maryland 20892.
Fordis
C M
CM
Howard
B H
BH
D000818:Animals; D003637:DEAE-Dextran / Q000494:pharmacology*; D004247:DNA / Q000494:pharmacology*; D003911:Dextrans / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000187:drug effects*
D003911:Dextrans; D004247:DNA; D003637:DEAE-Dextran