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Updated: Jan 20, 2026
Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
Published on: June 9, 2022
Establishment of an efficient reverse genetic system of Mumps virus S79 from cloned DNA
Duo Zhou1, Meng-Ying Zhu1, Yi-Long Wang2
1Zhejiang University School of Medicine, Hangzhou, China.
Background:
Mumps is a common type of respiratory infectious disease caused by mumps virus (MuV), and can be effectively prevented by vaccination. In this study, a reverse genetic system of MuV that can facilitate the rational design of safer, more efficient mumps vaccine candidates is established.
Methods:
MuV-S79 cDNA clone was assembled into a full-length plasmid by means of the GeneArt™ High-Order Genetic Assembly System, and was rescued via reverse genetic technology. RT-PCR, sequencing, and immunofluorescence assays were used for rMuV-S79 authentication. Viral replication kinetics and in vivo experimental models were used to evaluate the replication, safety, and immunogenicity of rMuV-S79.
Results:
A full-length cDNA clone of MuV-S79 in the assembly process was generated by a novel plasmid assemble strategy, and a robust reverse genetic system of MuV-S79 was successfully established. The established rMuV-S79 strain could reach a high virus titer in vitro. The average viral titer of rMuV-S79 in the lung tissues was 2.68 ± 0.14 log10PFU/g lung tissue, and rMuV-S79 group did not induce inflammation in the lung tissues in cotton rats. Neutralizing antibody titers induced by rMuV-S79 were high, long-lasting and could provide complete protection against MuV wild strain challenge.
Conclusion:
We have established a robust reverse genetic system of MuV-S79 which can facilitate the optimization of mumps vaccines. rMuV-S79 rescued could reach a high virus titer and the safety was proven in vivo. It could also provide complete protection against MuV wild strain challenge.
Insights
Researchers established a reverse genetic system for mumps virus (MuV) to develop improved mumps vaccines. The resulting rMuV-S79 strain showed high titers, safety in vivo, and provided complete protection against wild-type MuV challenge.
Area of Science:
- Virology
- Vaccinology
- Molecular Biology
Background:
- Mumps is a contagious respiratory illness caused by the mumps virus (MuV).
- Vaccination is an effective strategy for mumps prevention.
- Developing novel vaccine candidates requires advanced genetic tools.
Purpose of the Study:
- To establish a reverse genetic system for MuV.
- To facilitate the rational design of safer and more efficient mumps vaccine candidates.
Main Methods:
- Assembled a full-length MuV-S79 cDNA clone using a novel plasmid assembly strategy.
- Established a reverse genetic system for MuV rescue.
- Authenticated the rescued virus (rMuV-S79) using RT-PCR, sequencing, and immunofluorescence assays.
- Evaluated viral replication, safety, and immunogenicity in vitro and in vivo models.
Main Results:
- Successfully established a robust MuV-S79 reverse genetic system.
- The rescued rMuV-S79 strain achieved high titers in vitro.
- rMuV-S79 demonstrated safety in vivo, with no induced lung inflammation in cotton rats.
- rMuV-S79 induced high, long-lasting neutralizing antibody titers, conferring complete protection against wild-type MuV challenge.
Conclusions:
- A robust reverse genetic system for MuV-S79 was successfully established.
- The rescued rMuV-S79 strain exhibits high viral titers and proven in vivo safety.
- rMuV-S79 demonstrates potential as a vaccine candidate, offering complete protection against wild-type MuV.
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