Reverse genetics strategies for coronaviruses: platform construction and applications in vaccine development

Area of Science:

• Virology
• Genetics
• Global Health

Background:

• Novel coronaviruses emerge frequently, posing significant threats to global "One Health."
• High mutation rates and recombination events in coronaviruses, like Feline Coronavirus (FCoV-23) and SARS-CoV-2 variants, necessitate understanding their pathogenesis and cross-species transmission.
• Reverse genetics is crucial for functional virology but faces challenges with large coronavirus genomes and bacterial host instability.

Purpose of the Study:

• To systematically compare current reverse genetics strategies for coronavirus research.
• To critically evaluate the application of these platforms in understanding viral pathogenesis and developing next-generation vaccines.
• To identify solutions for technical bottlenecks in reverse genetics and design broad-spectrum countermeasures against emerging viral threats.

Main Methods:

• Systematic review and comparison of reverse genetics strategies.
• Evaluation of in vitro ligation, bacterial artificial chromosome (BAC) systems, and transformation-associated recombination (TAR) methods.
• Analysis of applications in deciphering pathogenic mechanisms and vaccine development.

Main Results:

• Comparison of different reverse genetics platforms across viral genera.
• Critical assessment of their utility in studying coronavirus pathogenesis and cross-species transmission.
• Identification of technical challenges and potential solutions for developing effective countermeasures.

Conclusions:

• Reverse genetics technologies are essential for advancing coronavirus research and developing countermeasures.
• Overcoming challenges in constructing reverse genetics systems is key to understanding and combating emerging viral threats.
• This review provides a framework for selecting and optimizing reverse genetics strategies for future research and vaccine development.