tetano
Editor, Senior Moderator
Proc Natl Acad Sci U S A
. 2021 Apr 13;118(15):e2025866118.
doi: 10.1073/pnas.2025866118.
Generation of SARS-CoV-2 reporter replicon for high-throughput antiviral screening and testing
Xi He[SUP] 1 [/SUP], Shuo Quan[SUP] 1 [/SUP], Min Xu[SUP] 1 [/SUP], Silveria Rodriguez[SUP] 1 [/SUP], Shih Lin Goh[SUP] 1 [/SUP], Jiajie Wei[SUP] 1 [/SUP], Arthur Fridman[SUP] 1 [/SUP], Kenneth A Koeplinger[SUP] 1 [/SUP], Steve S Carroll[SUP] 1 [/SUP], Jay A Grobler[SUP] 1 [/SUP], Amy S Espeseth[SUP] 1 [/SUP], David B Olsen[SUP] 1 [/SUP], Daria J Hazuda[SUP] 1 [/SUP], Dai Wang[SUP] 2 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) research and antiviral discovery are hampered by the lack of a cell-based virus replication system that can be readily adopted without biosafety level 3 (BSL-3) restrictions. Here, the construction of a noninfectious SARS-CoV-2 reporter replicon and its application in deciphering viral replication mechanisms and evaluating SARS-CoV-2 inhibitors are presented. The replicon genome is replication competent but does not produce progeny virions. Its replication can be inhibited by RdRp mutations or by known SARS-CoV-2 antiviral compounds. Using this system, a high-throughput antiviral assay has also been developed. Significant differences in potencies of several SARS-CoV-2 inhibitors in different cell lines were observed, which highlight the challenges of discovering antivirals capable of inhibiting viral replication in vivo and the importance of testing compounds in multiple cell culture models. The generation of a SARS-CoV-2 replicon provides a powerful platform to expand the global research effort to combat COVID-19.
Keywords: COVID-19; SARS-CoV-2; antivirals; high-throughput antiviral screening; replicon.
. 2021 Apr 13;118(15):e2025866118.
doi: 10.1073/pnas.2025866118.
Generation of SARS-CoV-2 reporter replicon for high-throughput antiviral screening and testing
Xi He[SUP] 1 [/SUP], Shuo Quan[SUP] 1 [/SUP], Min Xu[SUP] 1 [/SUP], Silveria Rodriguez[SUP] 1 [/SUP], Shih Lin Goh[SUP] 1 [/SUP], Jiajie Wei[SUP] 1 [/SUP], Arthur Fridman[SUP] 1 [/SUP], Kenneth A Koeplinger[SUP] 1 [/SUP], Steve S Carroll[SUP] 1 [/SUP], Jay A Grobler[SUP] 1 [/SUP], Amy S Espeseth[SUP] 1 [/SUP], David B Olsen[SUP] 1 [/SUP], Daria J Hazuda[SUP] 1 [/SUP], Dai Wang[SUP] 2 [/SUP]
Affiliations
- PMID: 33766889
- DOI: 10.1073/pnas.2025866118
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) research and antiviral discovery are hampered by the lack of a cell-based virus replication system that can be readily adopted without biosafety level 3 (BSL-3) restrictions. Here, the construction of a noninfectious SARS-CoV-2 reporter replicon and its application in deciphering viral replication mechanisms and evaluating SARS-CoV-2 inhibitors are presented. The replicon genome is replication competent but does not produce progeny virions. Its replication can be inhibited by RdRp mutations or by known SARS-CoV-2 antiviral compounds. Using this system, a high-throughput antiviral assay has also been developed. Significant differences in potencies of several SARS-CoV-2 inhibitors in different cell lines were observed, which highlight the challenges of discovering antivirals capable of inhibiting viral replication in vivo and the importance of testing compounds in multiple cell culture models. The generation of a SARS-CoV-2 replicon provides a powerful platform to expand the global research effort to combat COVID-19.
Keywords: COVID-19; SARS-CoV-2; antivirals; high-throughput antiviral screening; replicon.