Research progress and applications of reverse genetics systems for infectious bronchitis virus

Infectious bronchitis virus (IBV) poses a persistent threat to global poultry health, driving the need for advanced molecular tools to study and combat this pathogen. Reverse genetics has emerged as a pivotal technology in IBV research, enabling precise manipulation of the viral genome to investigate pathogenesis, design novel vaccines, and identify potential antiviral targets. This review systematically examines the development, applications, and challenges of reverse genetics platforms for IBV. Established methods, including vaccinia virus supported systems, in vitro ligation and transcription, targeted RNA recombination, bacterial artificial chromosome cloning, transformation associated recombination, and circular polymerase extension reaction are detailed, with their principles, advantages, and limitations highlighted. Furthermore, contributions of these platforms to elucidating gene function, rational vaccine design, and the development of IBV as a viral vector for multipathogen vaccines are discussed. Current technical hurdles, safety considerations, and knowledge gaps are addressed, along with future perspectives integrating CRISPR/Cas9, synthetic biology, and computational approaches. This comprehensive overview aims to guide researchers in selecting appropriate reverse genetics strategies and to inspire innovative solutions for IBV control.