Abstract
Coronaviruses (CoVs) are enveloped, single-stranded, positive-sense RNA viruses that cause respiratory, gastrointestinal, hepatic, and neurological diseases in humans and other animals. In recent years, frequent outbreaks of emerging and re-emerging CoVs have threatened animal and human health. However, an insufficient understanding of the mechanisms underlying CoV pathogenicity and cross-species transmission limits the development of drugs and vaccines against CoVs. Reverse genetic technology is a powerful tool for manipulating the genomes of CoVs and acquiring recombinant viruses, which allows researchers to better understand viral pathogenesis and develop genetically attenuated and marked vaccines and antiviral drugs. However, the large genomes of CoVs and the instability and toxicity of viral sequences in bacteria represent serious obstacles to the development of reverse genetic systems of CoVs. With the development of molecular biological methods, various new construction strategies have emerged. Accordingly, this review summarizes the construction strategies of CoV reverse genetics systems and their applications in studying pathogenesis, cross-species transmission, vaccine development, and drug screening, with the aim of providing an important reference for the prevention and control of CoVs.