title: Effects of G-gene Deletion and Replacement on Rabies Virus Vector Gene Expression.
abstract: The glycoprotein-gene (G gene) -deleted rabies virus (RV) vector is a powerful tool to examine the function and structure of neural circuits. We previously reported that the deletion of the G gene enhances the transgene expression level of the RV vector. However, the mechanism of this enhancement remains to be clarified. We presume that there are two possible factors for this enhancement. The first factor is the glycoprotein of RV, which shows cytotoxicity; thus, may cause a dysfunction in the translation process of infected cells. The second possible factor is the enhanced expression of the L gene, which encodes viral RNA polymerase. In the RV, it is known that the gene expression level is altered depending on the position of the gene. Since G-gene deletion displaces the L gene in the genome, the expression of the L gene and viral transcription may be enhanced. In this study, we compared the transgene expression level and viral transcription of three recombinant RV vectors. The effect of glycoprotein was examined by comparing the viral gene expression of G-gene-intact RV and G-gene-replaced RV. Despite the fact that the L-gene transcription level of these two RV vectors was similar, the G-gene-replaced RV vector showed higher viral transcription and transgene expression level than the G-gene-intact RV vector. To examine the effect of the position of the L gene, we compared the viral gene expression of the G-gene-deleted RV and G-gene-replaced RV. The G-gene-deleted RV vector showed higher L-gene transcription, viral transcription, and transgene expression level than the G-gene-replaced RV vector. These results indicate that G-gene deletion enhances the transgene expression level through at least two factors, the absence of glycoprotein and enhancement of L-gene expression. These findings enable investigators to design a useful viral vector that shows a controlled desirable transgene expression level in applications.
authors.affiliation: Division of Systems Neuroscience, Tohoku University Graduate School of Life Sciences, Sendai, Japan.
authors.forename: Sho
authors.initials: S
authors.lastname: Sato
authors.affiliation: Division of Systems Neuroscience, Tohoku University Graduate School of Life Sciences, Sendai, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Ohara
authors.affiliation: Division of Systems Neuroscience, Tohoku University Graduate School of Life Sciences, Sendai, Japan.
authors.forename: Ken-Ichiro
authors.initials: K
authors.lastname: Tsutsui
authors.affiliation: Division of Systems Neuroscience, Tohoku University Graduate School of Life Sciences, Sendai, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Iijima
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line / Q000821:virology; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006023:Glycoproteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011820:Rabies virus / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D019076:Transgenes; D014759:Viral Envelope Proteins / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D006023:Glycoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C064951:glycoprotein G, Rabies virus; C097625:L protein, Rabies virus; D012321:DNA-Directed RNA Polymerases