Abstract
Reverse genetics systems for viruses, the technology used to generate gene-engineered recombinant viruses from artificial genes, enable the study of the roles of the individual nucleotides and amino acids of viral genes and proteins in infectivity, replication, and pathogenicity. The successful development of a reverse genetics system for poliovirus in 1981 accelerated the establishment of protocols for other RNA viruses important for human health. Despite multiple efforts, rotavirus (RV), which causes severe gastroenteritis in infants, was refractory to reverse genetics analysis, and the first complete reverse genetics system for RV was established in 2017. This novel technique involves use of the fusogenic protein FAST (fusion-associated small transmembrane) derived from the bat-borne Nelson Bay orthoreovirus, which induces massive syncytium formation. Co-transfection of a FAST-expressing plasmid with complementary DNAs encoding RV genes enables rescue of recombinant RV. This review focuses on methodological insights into the reverse genetics system for RV and discusses applications and potential improvements to this system.
Highlights
Advances in molecular biology, biochemistry, and genetics have contributed to our understanding of the basic biology of viruses
The principles of reverse genetics systems are based on the viral replication cycle: Once the DNA or RNA corresponding to the viral genomes is introduced into cells by plasmid vectors, viral vectors, or direct transfection, the viral genome and proteins are synthesized, resulting in the production of recombinant virus
Viral proteins can be synthesized from the viral messenger RNA template by the cellular translation machinery, indicating that the minimum requirement for the RV replication cycle is the complete set of 5 -capped (+)single-stranded RNA (ssRNA) transcribed from viral double-stranded RNA (dsRNA)
Summary
Biochemistry, and genetics have contributed to our understanding of the basic biology of viruses. Reverse genetics technology, which can yield geneengineered recombinant viruses from artificial genomes, has enabled studies of the function of viral proteins, molecular mechanisms of viral pathogenesis, viral vectors, and vaccine development. The family Reoviridae includes human and animal pathogens such as rotavirus (RV), mammalian orthoreovirus (MRV), Nelson Bay orthoreovirus (NBV), and bluetongue virus (BTV). MRV infects the respiratory and enteric tracts but is rarely associated with disease [8] It selectively infects and kills cancer cells and has been developed as an oncolytic viral therapy [9]. Due to the complex structure of their genomes, the development of a helper virus–independent, complete reverse genetics system of the family Reoviridae lagged behind that of other virus families associated with human and animal health. This review focuses on technical insights and applications of the reverse genetics system for RV
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
