PMID:24244660

Citation	Ohara, S, Sato, S, Oyama, K, Tsutsui, K and Iijima, T (2013) Rabies virus vector transgene expression level and cytotoxicity improvement induced by deletion of glycoprotein gene. PLoS ONE 8:e80245
Abstract	The glycoprotein (G) of rabies virus (RV) is required for binding to neuronal receptors and for viral entry. G-deleted RV vector is a powerful tool for investigating the organization and function of the neural circuits. It gives the investigator the ability to genetically target initial infection to particular neurons and to control trans-synaptic propagation. In this study we have quantitatively evaluated the effect of G gene deletion on the cytotoxicity and transgene expression level of the RV vector. We compared the characteristics of the propagation-competent RV vector (rHEP5.0-CVSG-mRFP) and the G-deleted RV vector (rHEP5.0-ΔG-mRFP), both of which are based on the attenuated HEP-Flury strain and express monomeric red fluorescent protein (mRFP) as a transgene. rHEP5.0-ΔG-mRFP showed lower cytotoxicity than rHEP5.0-CVSG-mRFP, and within 16 days of infection we found no change in the basic electrophysiological properties of neurons infected with the rHEP5.0-ΔG-mRFP. The mRFP expression level of rHEP5.0-ΔG-mRFP was much higher than that of rHEP5.0-CVSG-mRFP, and 3 days after infection the retrogradely infected neurons were clearly visualized by the expressed fluorescent protein without any staining. This may be due to the low cytotoxicity and/or the presumed change in the polymerase gene (L) expression level of the G-deleted RV vector. Although the mechanisms remains to be clarified, the results of this study indicate that deletion of the G gene greatly improves the usability of the RV vector for studying the organization and function of the neural circuits by decreasing the cytotoxicity and increasing the transgene expression level.
Links	PubMed PMC3820615 Online version:10.1371/journal.pone.0080245
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9RHAB:Q85422	part_of	GO:0019031: viral envelope	ECO:0000315: mutant phenotype evidence used in manual assertion		C		Seeded From UniProt	complete
9RHAB:Q85422		GO:0019031: viral envelope	ECO:0000315:		C		Figure 1B and 1C show how the g protein of the rabies virus is required for infecting cells via binding to neuronal receptors and for viral entry. When the g protein was knocked out of the RV vector, rHEP5.0-delta G, the number of infected cells failed to increase whereas the propagation-competent RV vector rHEP5.0-CVSG was unhindered by infecting cells because it had a functional g protein.	complete CACAO 9735

References

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