Post-transcriptional regulatory element boosts baculovirus-mediated gene expression in vertebrate cells.
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.
Mähönen
Anssi J
AJ
A.I. Virtanen Institute, Department of Biotechnology and Molecular Medicine, Kuopio, Finland.
Airenne
Kari J
KJ
Purola
Sanna
S
Peltomaa
Erik
E
Kaikkonen
Minna U
MU
Riekkinen
Mervi S
MS
Heikura
Tommi
T
Kinnunen
Kati
K
Roschier
Miia M
MM
Wirth
Thomas
T
Ylä-Herttuala
Seppo
S
D000818:Animals; D016367:Baculoviridae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D002087:Butyrates / Q000494:pharmacology; D002460:Cell Line; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D017919:Hepatitis B Virus, Woodchuck; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics*; D014714:Vertebrates; D001616:beta-Galactosidase / Q000378:metabolism
D002087:Butyrates; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase