Truncated vesicular stomatitis virus G protein improves baculovirus transduction efficiency in vitro and in vivo.
Pseudotyping of viral vectors has been widely used to enhance viral transduction efficiency. One of the most popular pseudotyping proteins has been the G-protein of the vesicular stomatitis virus, VSV-G. In the present study, we show that the 21-amino-acid ectodomain with transmembrane and cytoplasmic tail domains of VSV-G (VSV-GED) augments baculovirus-mediated gene delivery in vertebrate cells by aiding viral entry. The VSV-GED pseudotyped virus replicated efficiently in insect cells yielding high titers. Five out of six studied cell lines showed improved transduction, as measured by a number of transduced cells or transgene expression level. Nearly 15-fold increase in the transduction efficiency was detected in rat malignant glioma cells as compared to the control virus. In the rat brain, transgene expression could be detected in the walls of lateral ventricles and in subarachnoid membranes. Increased transduction efficiency was also observed in the rabbit muscle. Our results suggest that VSV-GED enhances baculoviral gene transfer by augmenting gp64-mediated endosomal release. Moreover, no cytotoxicity was associated with improved gene transfer efficiency. Thus, VSV-GED pseudotyping provides a simple means to enhance baculovirus-mediated gene transfer in vitro and in vivo.
Kaikkonen
M U
MU
AI Virtanen Institute, Department of Biotechnology and Molecular Medicine, University of Kuopio, Kuopio, Finland.
Räty
J K
JK
Airenne
K J
KJ
Wirth
T
T
Heikura
T
T
Ylä-Herttuala
S
S
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001921:Brain / Q000378:metabolism; D002478:Cells, Cultured; D011992:Endosomes / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D005910:Glioma / Q000628:therapy*; D015151:Immunoblotting; D008562:Membrane Glycoproteins / Q000009:adverse effects / Q000235:genetics* / Q000378:metabolism; D023421:Models, Animal; D018482:Muscle, Skeletal / Q000378:metabolism; D009380:Neoplasms, Nerve Tissue / Q000628:therapy*; D017434:Protein Structure, Tertiary; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D014161:Transduction, Genetic / Q000379:methods*; D014759:Viral Envelope Proteins / Q000009:adverse effects / Q000235:genetics* / Q000378:metabolism
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins