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J Biotechnol. 2007 Aug 1;131(1):1-8. Epub 2007 May 31.

Post-transcriptional regulatory element boosts baculovirus-mediated gene expression in vertebrate cells.

Author information

1
A.I. Virtanen Institute, Department of Biotechnology and Molecular Medicine, Kuopio, Finland.

Abstract

Baculoviruses can express transgenes in a wide range of vertebrate cells. However, in some cells transgene expression is weak. To enhance transgene expression, we studied the effect of the Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) on baculovirus (BV)-mediated gene expression of several transgenes. A significant increase in BV-mediated gene expression was detected in several cell lines. A 10-fold increase in transgene expression was observed with the WPRE as determined by the percentage of positive cells and mean fluorescence intensity (MFI). Furthermore, a combination of optimized cell culture medium and WPRE virus led to more than a 60-fold increase in gene expression. In accordance, elevated mRNA and protein levels were detected in WPRE-virus transduced cells. In HepG2 and RaaSMC, WPRE-mediated enhancement was comparable to the previously shown positive effect of sodium butyrate on BV-mediated gene expression. Thus, inclusion of the WPRE into a baculovirus vector provides a simple means to improve BV-mediated gene expression in vertebrate cells.

PMID:
17617485
DOI:
10.1016/j.jbiotec.2007.05.022
[Indexed for MEDLINE]

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