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PMID: 3286163 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Short-term efficient expression of transfected DNA in human hematopoietic cells by electroporation: definition of parameters and use of chemical stimulators.

DNA (Mary Ann Liebert, Inc.) ·Vol. 7 ·No. 3 ·1988-04-00 ·Pages 203-9

Satyabhama S, Epstein AL

Abstract

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.

MeSH Terms
Acetyltransferases/genetics Cell Line Chloramphenicol O-Acetyltransferase DNA/genetics Genes Genes, Bacterial Genetic Techniques Hematopoietic Stem Cells/metabolism Humans Leukemia, Erythroblastic, Acute Transcription, Genetic Transfection
Chemicals
DNA Acetyltransferases Chloramphenicol O-Acetyltransferase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Satyabhama S
Department of Pathology, USC Medical Center, Los Angeles 90033.
Epstein A L
Article Info
Journal
DNA (Mary Ann Liebert, Inc.)
Abbr.
DNA
ISSN
0198-0238
Published
1988-04-00
Pages
203-9
Language
English
Region
United States
NLM ID
8302432
Subset
IM
Grants
NCI NIH HHS · R01-CA40608 · United States
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