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Optimization of short‐term transgene expression by sodium butyrate and ubiquitous chromatin opening elements (UCOEs)
Author(s) -
de Poorter Jolanda J.,
Lipinski Kai S.,
Nelissen Rob G. H. H.,
Huizinga Tom W. J.,
Hoeben Rob C.
Publication year - 2007
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1057
Subject(s) - sodium butyrate , transgene , genetic enhancement , microbiology and biotechnology , reporter gene , viral vector , gene expression , biology , luciferase , chromatin , cell culture , electroporation , gene , cancer research , transfection , recombinant dna , biochemistry , genetics
Background Predictable and adequate transgene expression is essential for clinical gene therapy. Several studies have focused on optimization of transgene expression. In this study the effect of sodium butyrate (NaB) and a ubiquitous chromatin opening element (UCOE) on short‐term gene expression after adenovirus‐mediated gene transfer in fibroblastic interface cells from periprosthetic tissue in loosened orthopedic implants is investigated. Methods Cultures of diploid human interface cells from four patients were infected with an adenovirus type‐5 vector that carries the luciferase gene driven by the cytomegalovirus (CMV) promoter as a reporter. In addition, viruses with a UCOE were evaluated. Twenty‐four hours after infection NaB was added in concentrations of 0 to 9 mM. Luciferase activity was tested after a further 24 h. Results NaB in a concentration of 6 mM caused a 7‐ to 16‐fold increase in reporter gene expression compared to control condition. There was no difference in reporter gene expression when cells were infected with Ad.1.5UCOE‐CMV.Luc compared to Ad.CMV.Luc. A combination of NaB and a UCOE had no advantage over NaB alone. Conclusions Addition of NaB results in a marked increase in transgene expression in cultured cells. This would allow the enhancement of the expression of the transgene, without requiring a higher vector dose. Butyrate administration could not be substituted by inclusion of UCOEs in the vector. It remains to be established whether the effective concentrations of butyrate can be obtained in vivo . Copyright © 2007 John Wiley & Sons, Ltd.