Glycoprotein gene relocation in rabies virus.

Area of Science:

• Virology
• Molecular Biology
• Vaccinology

Background:

• Relocating genes in rabies virus using reverse genetics has been challenging.
• Understanding gene function and virus pathogenicity requires manipulation of viral genomes.

Purpose of the Study:

• To develop an efficient reverse genetics system for rabies virus (Evelyn-Rokitnicki-Abelseth strain).
• To create a reshuffled rabies virus with an altered gene order (ERAggm) by switching the glycoprotein and matrix protein genes.
• To assess the replication efficiency and potential applications of the novel ERAgm virus.

Main Methods:

• Utilized reverse genetics to engineer the Evelyn-Rokitnicki-Abelseth (ERA) rabies virus.
• Implemented an autogene plasmid and T7 RNA polymerase with a nuclear location signal for virus recovery.
• Rearranged the viral genome to create ERAgm (N-P-G-M-L gene order) by swapping the glycoprotein (G) and matrix protein (M) genes.

Main Results:

• Successfully generated the reshuffled ERAgm rabies virus.
• ERAggm replicated efficiently, achieving titers comparable to the parental ERA virus (N-P-M-G-L gene order) in BSR cells (10^9 ffu/ml).
• The altered gene order did not impede viral replication.

Conclusions:

• The developed reverse genetics system enables efficient manipulation of the rabies virus genome.
• The reshuffled ERAgm virus provides a new platform for investigating rabies virus pathogenicity.
• This approach offers a potential novel strategy for developing improved rabies vaccines.