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Measuring Alphavirus Fidelity Using Non-Infectious Virus Particles.
Edward I Patterson1,2, Kamil Khanipov3, Daniele M Swetnam4
1Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Viruses
|May 21, 2020
Summary
Altering viral mutation rates can create live-attenuated vaccines. Researchers deleted a VEEV cleavage site to study mutations without selection, revealing lower mutation frequencies in engineered viruses.
Area of Science:
- Virology
- Vaccine Development
- Molecular Biology
Background:
- RNA viruses incorporate mutations at an optimal frequency, crucial for live-attenuated vaccine strategies.
- Assessing the impact of specific mutations on viral fidelity has been challenging due to rapid selection during replication.
Purpose of the Study:
- To develop a method for evaluating the effect of mutations on viral mutation frequency without the confounding factor of selection.
- To investigate the role of the PE2 cleavage site in Venezuelan equine encephalitis virus (VEEV) fidelity.
Main Methods:
- Deletion of structural polyprotein PE2 cleavage site residues (E3D56-59) in VEEV TC-83 to create non-infectious particles.
- Utilizing next-generation sequencing to analyze mutation frequencies in engineered VEEV particles.
Main Results:
- Deletion of the PE2 cleavage site rendered VEEV non-infectious, enabling mutation analysis without selection.
- Fidelity mutants exhibited significantly lower transversion and overall mutation frequencies compared to the control VEEV TC-83 E3D56-59.
Conclusions:
- The PE2 cleavage site deletion strategy effectively isolates the effect of mutations on viral fidelity.
- This method provides a platform for evaluating fidelity mutants across medically important alphaviruses.
