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

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• 作者：Anssi J. Mä ; hö ; nen ; Kari J. Airenne ; Sanna Purola ; Erik Peltomaa ; Minna U. Kaikkonen ; Mervi S. Riekkinen ; Tommi Heikura ; Kati Kinnunen ; Miia M. Roschier ; Thomas Wirth ; Seppo Ylä ; -Herttual
• 关键词：Baculovirus ; Gene delivery ; Post-transcriptional element ; Gene therapy ; WPRE ; Transduction
• 刊名：Journal of Biotechnology
• 出版年：2007
• 出版时间：1 August 2007
• 年：2007
• 卷：131
• 期：1
• 页码：1-8
• 全文大小：1211 K

文摘

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.