PT  - JOURNAL ARTICLE
AU  - Yang Zhang
AU  - Wuhui Song
AU  - Shuiye Chen
AU  - Zhenghong Yuan
AU  - Zhigang Yi
TI  - A bacterial artificial chromosome (BAC)-vectored noninfectious replicon of SARS-CoV-2
AID  - 10.1101/2020.09.11.294330
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.09.11.294330
4099  - http://biorxiv.org/content/early/2020/09/12/2020.09.11.294330.short
4100  - http://biorxiv.org/content/early/2020/09/12/2020.09.11.294330.full
AB  - Vaccines and antiviral agents are in urgent need to stop the COVID-19 pandemic. To facilitate antiviral screening against SARS-CoV-2 without requirement for high biosafety level facility, we developed a bacterial artificial chromosome (BAC)-vectored replicon of SARS-CoV-2, nCoV-SH01 strain, in which secreted Gaussia luciferase (sGluc) was encoded in viral subgenomic mRNA as a reporter gene. The replicon was devoid of structural genes spike (S), membrane (M), and envelope (E). Upon transfection, the replicon RNA replicated in various cell lines, and was sensitive to interferon alpha (IFN-α), remdesivir, but was resistant to hepatitis C virus inhibitors daclatasvir and sofosbuvir. Replication of the replicon was also sensitive overexpression of zinc-finger antiviral protein (ZAP). We also constructed a four-plasmid in-vitro ligation system that is compatible with the BAC system, which makes it easy to introduce desired mutations into the assembly plasmids for in-vitro ligation. This replicon system would be helpful for performing antiviral screening and dissecting virus-host interactions.Competing Interest StatementThe authors have declared no competing interest.