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A Fosmid-Based System for the Generation of Recombinant Cercopithecine Alphaherpesvirus 2 Encoding Reporter Genes
Ekaterina Chukhno1, Sabine Gärtner1, Abdul Rahman Siregar1,2
1Infection Biology Unit, German Primate Center-Leibniz Institute for Primate Research, 37077 Göttingen, Germany.
Abstract:
The transmission of Macacine alphaherpesvirus 1 (McHV-1) from macaques, the natural host, to humans causes encephalitis. In contrast, human infection with Cercopithecine alphaherpesvirus 2 (CeHV-2), a closely related alphaherpesvirus from African vervet monkeys and baboons, has not been reported and it is believed that CeHV-2 is apathogenic in humans. The reasons for the differential neurovirulence of McHV-1 and CeHV-2 have not been explored on a molecular level, in part due to the absence of systems for the production of recombinant viruses. Here, we report the generation of a fosmid-based system for rescue of recombinant CeHV-2. Moreover, we show that, in this system, recombineering can be used to equip CeHV-2 with reporter genes. The recombinant CeHV-2 viruses replicated with the same efficiency as uncloned, wt virus and allowed the identification of cell lines that are highly susceptible to CeHV-2 infection. Collectively, we report a system that allows rescue and genetic modification of CeHV-2 and likely other alphaherpesviruses. This system should aid future analysis of CeHV-2 biology.
Insights
Researchers developed a new system to genetically modify Cercopithecine alphaherpesvirus 2 (CeHV-2). This tool enables the study of CeHV-2, a virus believed to be non-pathogenic in humans, unlike its relative McHV-1.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Macacine alphaherpesvirus 1 (McHV-1) causes encephalitis in humans, while closely related Cercopithecine alphaherpesvirus 2 (CeHV-2) is considered apathogenic.
- The molecular basis for this differential neurovirulence is unknown due to a lack of tools for studying CeHV-2.
Purpose of the Study:
- To develop a system for the rescue and genetic manipulation of CeHV-2.
- To enable future investigations into the biology and pathogenicity of CeHV-2.
Main Methods:
- Generation of a fosmid-based system for CeHV-2 rescue.
- Utilizing recombineering to insert reporter genes into CeHV-2.
- Characterizing the replication efficiency of recombinant CeHV-2.
Main Results:
- Successfully generated a system for rescuing and genetically modifying CeHV-2.
- Recombinant CeHV-2 viruses replicated efficiently, comparable to wild-type virus.
- Identified cell lines highly susceptible to CeHV-2 infection.
Conclusions:
- A novel system for CeHV-2 rescue and genetic modification has been established.
- This system facilitates the study of CeHV-2 replication and host cell interactions.
- The developed system is potentially applicable to other alphaherpesviruses.
