A series of SARS-CoV-2 sub-genomic replicons as artificial bacterial chromosomes for the screening of SARS-CoV-2 in studies such as drug discovery and inhibitors assay.
Coronavirus disease (COVID-19) is a deadly infectious disease caused by the new SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) virus. SARS-CoV-2 is a beta coronavirus, similar to MERS-CoV and SARS-CoV, and it is the cause of the current pandemic. COVID-19 typically presents as a respiratory illness with symptoms including cough, fever, and difficulty breathing. Because COVID-19 is highly contagious, screening and contact tracing methods are of the utmost importance in stopping the spread of this disease.
Emory researchers have generated multiple replicons of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as bacterial artificial chromosomes (BACs). Replicons are self-replicative systems in which the genes encoding viral structural proteins are replaced by a reporter gene, such as a fluorescent protein to allow for the visual detection of RNA replication. Transfection of susceptible cells with replicon systems results in the transient expression of the reporter protein, whose level of activity then reflects the extent of viral replication for a limited period of time. The SARS-CoV-2 replicon system is a valuable tool that can be used for high throughput screening and characterization of SARS-CoV-2 replicase inhibitors at a lower biosafety level. Specifically, replicons were engineered using bacterial artificial chromosomes (BACs), and ligation of several cDNA fragments, that were either purchased or prepared by PCR from infectious virus genomes.
In vitro assays, pre-clinical data only.
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