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Attenuated MuV-S79 as vector stably expressing foreign gene
Duo Zhou1, Meng-Ying Zhu1, Yi-Long Wang2
1Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
Background:
To describe mumps virus (MuV) used as a vector to express enhanced green fluorescent protein (EGFP) or red fluorescent protein (RFP) genes.
Methods:
Molecular cloning technique was applied to establish the cDNA clones of recombinant mumps viruses (rMuVs). rMuVs were recovered based on our reverse genetic system of MuV-S79. The properties of rMuVs were determined by growth curve, plaque assay, fluorescent microscopy and determination of fluorescent intensity.
Results:
Three recombinant viruses replicated well in Vero cells and similarly as parental rMuV-S79, expressed heterologous genes in high levels, and were genetically stable in at least 15 passages.
Conclusion:
rMuV-S79 is a promising platform to accommodate foreign genes like marker genes, other antigens and immunomodulators for addressing various diseases.
Insights
Recombinant mumps viruses (rMuVs) carrying fluorescent genes were successfully created and demonstrated stable replication and high gene expression in cell cultures. This indicates their potential as versatile gene delivery vectors for various applications.
Area of Science:
- Virology
- Molecular Biology
- Gene Therapy
Background:
- Mumps virus (MuV) is explored as a potential vector for expressing foreign genes.
- Focus on utilizing MuV to express enhanced green fluorescent protein (EGFP) and red fluorescent protein (RFP).
Purpose of the Study:
- To develop and characterize recombinant mumps viruses (rMuVs) capable of expressing fluorescent marker genes.
- To assess the feasibility of using the MuV-S79 reverse genetics system for gene expression.
Main Methods:
- Molecular cloning techniques were employed to create cDNA clones of rMuVs.
- Recombinant viruses were recovered using the MuV-S79 reverse genetic system.
- Virus properties were evaluated through growth curves, plaque assays, and fluorescent microscopy.
Main Results:
- Three rMuVs replicated efficiently in Vero cells, comparable to the parental virus.
- High-level expression of heterologous genes (EGFP/RFP) was achieved.
- Genetic stability was confirmed for at least 15 passages.
Conclusions:
- The rMuV-S79 platform is a promising tool for accommodating foreign genes, including marker genes, antigens, and immunomodulators.
- This platform holds potential for applications in addressing various diseases.
- rMuVs demonstrate robust gene expression and stability, supporting their use as gene delivery vectors.
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