Short-term efficient expression of transfected DNA in human hematopoietic cells by electroporation: definition of parameters and use of chemical stimulators.
The efficiency of DNA transfer into human hematopoietic cells by electroporation was investigated and compared to conventional transfection procedures. Important parameters of electroporation were optimized in human erythroleukemia cells using the chloramphenicol acetyltransferase (acetyl-CoA; chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28) gene linked to the cytomegaloviral enhancer-promoter. In addition, selected chemicals with different modes of action were studied for their ability to aid DNA entry and gene expression in this system, and several were found to enhance gene transfection by electroporation in a significant manner. Using these chemical stimulators, many but not all human and mouse suspension cultures tested were successfully electroporated by the Baekon 2000 instrument. From these studies, it appears that electroporation can be enhanced by chemical additives. Because of its efficiency, reproductivity, and convenience electroporation is an attractive method of gene transfer in human hematopoietic cells.
Satyabhama
S
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Department of Pathology, USC Medical Center, Los Angeles 90033.
Epstein
A L
AL
D000123:Acetyltransferases / Q000235:genetics; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase; D004247:DNA / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005821:Genetic Techniques; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute; D014158:Transcription, Genetic; D014162:Transfection
D004247:DNA; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase