The production and application of SARS-CoV-2 reverse genetic systems to facilitate vaccine development and biosafe drug discovery platforms.

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The development of vaccines and antiviral drugs against SARS-CoV-2 is hampered by the requirement to grow the virus under high-containment, which is limited to few laboratories worldwide. We have recently established a rapid procedure to produce a DNA copy of the SARS-CoV-2 RNA genome, that can be used to genetically engineer new viruses via a process termed "reverse genetics". We now propose to engineer the SARS-CoV-2 genome to produce 1) a SARS-CoV-2 virus that expresses a green fluorescent protein (turboGFP) and 2) a disabled biosafe form of the virus (replicon) that expresses turboGFP instead of the structurally important spike protein. When introduced into suitable cells, the replicon reproduces continously but does not produce infectious virus particles. The two engineered forms of the virus can then be used to develop high-throughput fluorescence based assays to monitor the entry of the virus into cells and its replication. The first can be used to identify antibodies induced by vaccine candidates that neutralise virus infectivity, and also viral mutations that allow escape from the antibody response. The replicon can be used for rapid screening of antiviral compounds, at a lower containment level, which would allow many more laboratories to participate in drug discovery efforts.