| Literature DB >> 17617485 |
Anssi J Mähönen1, Kari J Airenne, Sanna Purola, Erik Peltomaa, Minna U Kaikkonen, Mervi S Riekkinen, Tommi Heikura, Kati Kinnunen, Miia M Roschier, Thomas Wirth, Seppo Ylä-Herttuala.
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.Entities:
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Year: 2007 PMID: 17617485 DOI: 10.1016/j.jbiotec.2007.05.022
Source DB: PubMed Journal: J Biotechnol ISSN: 0168-1656 Impact factor: 3.307