# Research Progress and Applications of the Rotavirus Reverse Genetics System

## TL;DR

This review discusses how advances in rotavirus reverse genetics have enabled precise genetic engineering of the virus, aiding vaccine development and antiviral research.

## Contribution

The paper outlines the evolution and current capabilities of rotavirus reverse genetics systems, emphasizing recent breakthroughs and applications.

## Key findings

- The first fully plasmid-based reverse genetics system was established in 2017, eliminating the need for helper viruses.
- Optimizations like codon modification and CRISPR/Cas9 integration have improved system efficiency for diverse strains.
- Reverse genetics now supports vaccine design, antiviral screening, and understanding cross-species transmission.

## Abstract

Rotavirus (RV) remains a leading cause of severe, dehydrating diarrhea in infants and young animals, imposing significant global morbidity, mortality, and economic burdens. Historically, the development of effective interventions has been impeded by the technical challenges inherent to manipulating its complex, segmented RNA genome. This review highlights pivotal advancements in rotavirus reverse genetics—a technology enabling the de novo generation of infectious virus entirely from cloned cDNA. Transitioning from early, inefficient systems, this technology has matured into a viable and versatile platform, now facilitating the precise genetic engineering of diverse rotavirus strains from both human and animal hosts. These capabilities are instrumental for elucidating viral pathogenesis, designing next-generation vaccines, establishing high-throughput platforms for antiviral drug discovery, and investigating transmission dynamics. Collectively, these advances provide essential tools to accelerate the development of improved interventions to mitigate the global burden of rotavirus disease.

Rotavirus is a leading cause of severe, dehydrating diarrhea in infants and young animals, causing significant global morbidity and mortality. For decades, research was hindered by challenges in establishing reverse genetics systems due to the virus’s complex segmented genome and poor cell culture adaptation. The first helper virus-dependent system (2006) was limited by low efficiency. A 2017 breakthrough established the first fully plasmid-based system, which eliminated helper viruses and revolutionized the field. Subsequent optimizations, such as codon modification and CRISPR/Cas9 integration, have significantly enhanced efficiency, enabling viable systems for diverse human and animal strains. This narrative review summarizes the evolution and technological milestones of rotavirus reverse genetics. We discuss critical applications in analyzing viral gene function, developing novel vaccines, screening antiviral drugs, and investigating cross-species transmission. Finally, we provide an outlook on the future prospects of this transformative technology.