Reprogramming Rotavirus: Reverse Genetics-Driven Design of Viral Vector Platforms

Rotaviruses remain the leading cause of severe dehydrating diarrhea and associated mortality in children under five years of age worldwide. The successful global rollout of live-attenuated oral rotavirus vaccines has dramatically reduced rotavirus gastroenteritis morbidity and mortality, unequivocally demonstrating their excellent safety profile and potent induction of mucosal immunity. These attributes highlight the substantial potential of rotaviruses as novel oral mucosal vaccine vectors. Recent breakthroughs in reverse genetics, particularly the establishment of a fully plasmid-based system in 2017, have enabled precise insertion and stable expression of foreign antigens at targeted genomic loci. This advance has opened a viable pathway for developing multivalent oral mucosal vaccines. This review traces the historical development of rotavirus reverse genetics and summarizes the latest progress in its application as a vaccine vector platform. We focus on key strategies for foreign gene insertion and immunological outcomes in animal models, while critically evaluating persistent challenges—virus rescue efficiency, genetic stability of inserts, and limitations of current animal models—and outlining the rational design framework for RV-based vectors with improved stability, expression efficiency, and immunogenicity.