A Fluorescent Reporter Virus Toolkit for Interrogating Enterovirus Biology and Host Interactions

Area of Science:

• Virology
• Molecular Biology
• Pathogen Research

Background:

• Enteroviruses are common human pathogens causing diverse illnesses.
• Understanding enterovirus biology is crucial for developing vaccines and antivirals.
• Fluorescent reporter viruses aid in studying viral behavior and host interactions.

Purpose of the Study:

• To identify optimal sites for fluorescent reporter gene insertion in coxsackievirus B3.
• To establish experimental workflows for analyzing viral fitness, neutralization, and antiviral efficacy.
• To enable broader application of reporter virus technology for enterovirus research.

Main Methods:

• Constructed recombinant coxsackievirus B3 with fluorescent reporters at three distinct polyprotein sites.
• Assessed viral fitness and replication efficiency for each reporter insertion site.
• Developed and validated assays for viral fitness, serum neutralization, antiviral efficacy, and recombination.

Main Results:

• Identified a specific site within the viral polyprotein that tolerates reporter gene insertion with minimal impact on viral fitness.
• Demonstrated the utility of fluorescent reporter viruses in quantitative assays for viral characteristics.
• Established robust workflows applicable to various enterovirus studies.

Conclusions:

• Reporter gene insertion site significantly impacts enterovirus fitness; a specific site is optimal.
• Fluorescent reporter viruses provide a versatile platform for advancing enterovirus research, vaccine development, and antiviral screening.
• The developed methods are adaptable for studying other medically relevant enteroviruses.