Screening of SARS-CoV-2 antivirals through a cell-based RNA-dependent RNA polymerase (RdRp) reporter assay
COVID-19 (Coronavirus Disease 2019) caused by SARS-CoV-2 (Severe Acute Respiratory Syndrome CoronaVirus-2) continues to pose an international public health threat and thus far, has resulted in greater than 6.4 million deaths worldwide. Vaccines are critical tools to limit COVID-19 spread, but antivi...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-08-01
|
| Series: | Cell Insight |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2772892722000438 |
| _version_ | 1798026964246724608 |
|---|---|
| author | Timsy Uppal Kai Tuffo Svetlana Khaiboullina Sivani Reganti Mark Pandori Subhash C. Verma |
| author_facet | Timsy Uppal Kai Tuffo Svetlana Khaiboullina Sivani Reganti Mark Pandori Subhash C. Verma |
| author_sort | Timsy Uppal |
| collection | DOAJ |
| description | COVID-19 (Coronavirus Disease 2019) caused by SARS-CoV-2 (Severe Acute Respiratory Syndrome CoronaVirus-2) continues to pose an international public health threat and thus far, has resulted in greater than 6.4 million deaths worldwide. Vaccines are critical tools to limit COVID-19 spread, but antiviral drug development is an ongoing global priority due to fast-spreading COVID-19 variants that may elude vaccine efficacies. The RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2 is an essential enzyme of viral replication and transcription machinery complex. Therefore, the RdRp is an attractive target for the development of effective anti-COVID-19 therapeutics. In this study, we developed a cell-based assay to determine the enzymatic activity of SARS-CoV-2 RdRp through a luciferase reporter system. The SARS-CoV-2 RdRp reporter assay was validated using known inhibitors of RdRp polymerase, remdesivir along with other anti-virals including ribavirin, penciclovir, rhoifolin, 5′CT, and dasabuvir. Dasabuvir (an FDA-approved drug) exhibited promising RdRp inhibitory activity among these inhibitors. Anti-viral activity of dasabuvir was also tested on the replication of SARS-CoV-2 through infection of Vero E6 cells. Dasabuvir inhibited the replication of SARS-CoV-2, USA-WA1/2020 as well as B.1.617.2 (delta variant) in Vero E6 cells in a dose-dependent manner with EC50 values 9.47 μM and 10.48 μM, for USA-WA1/2020 and B.1.617.2 variants, respectively. Our results suggest that dasabuvir can be further evaluated as a therapeutic drug for COVID-19. Importantly, this system provides a robust, target-specific, and high-throughput screening compatible (z- and z’-factors of >0.5) platforms that will be a valuable tool for screening SARS-CoV-2 RdRp inhibitors. |
| first_indexed | 2024-04-11T18:43:53Z |
| format | Article |
| id | doaj.art-24ec9cd407bd498c81bcad367080804c |
| institution | Directory Open Access Journal |
| issn | 2772-8927 |
| language | English |
| last_indexed | 2024-04-11T18:43:53Z |
| publishDate | 2022-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Insight |
| spelling | doaj.art-24ec9cd407bd498c81bcad367080804c2022-12-22T04:08:53ZengElsevierCell Insight2772-89272022-08-0114100046Screening of SARS-CoV-2 antivirals through a cell-based RNA-dependent RNA polymerase (RdRp) reporter assayTimsy Uppal0Kai Tuffo1Svetlana Khaiboullina2Sivani Reganti3Mark Pandori4Subhash C. Verma5Department of Microbiology and Immunology, University of Nevada, Reno, NV, 89557, USADepartment of Microbiology and Immunology, University of Nevada, Reno, NV, 89557, USADepartment of Microbiology and Immunology, University of Nevada, Reno, NV, 89557, USADepartment of Microbiology and Immunology, University of Nevada, Reno, NV, 89557, USANevada State Public Health Laboratory, Reno, NV, 89557, USADepartment of Microbiology and Immunology, University of Nevada, Reno, NV, 89557, USA; Corresponding author.COVID-19 (Coronavirus Disease 2019) caused by SARS-CoV-2 (Severe Acute Respiratory Syndrome CoronaVirus-2) continues to pose an international public health threat and thus far, has resulted in greater than 6.4 million deaths worldwide. Vaccines are critical tools to limit COVID-19 spread, but antiviral drug development is an ongoing global priority due to fast-spreading COVID-19 variants that may elude vaccine efficacies. The RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2 is an essential enzyme of viral replication and transcription machinery complex. Therefore, the RdRp is an attractive target for the development of effective anti-COVID-19 therapeutics. In this study, we developed a cell-based assay to determine the enzymatic activity of SARS-CoV-2 RdRp through a luciferase reporter system. The SARS-CoV-2 RdRp reporter assay was validated using known inhibitors of RdRp polymerase, remdesivir along with other anti-virals including ribavirin, penciclovir, rhoifolin, 5′CT, and dasabuvir. Dasabuvir (an FDA-approved drug) exhibited promising RdRp inhibitory activity among these inhibitors. Anti-viral activity of dasabuvir was also tested on the replication of SARS-CoV-2 through infection of Vero E6 cells. Dasabuvir inhibited the replication of SARS-CoV-2, USA-WA1/2020 as well as B.1.617.2 (delta variant) in Vero E6 cells in a dose-dependent manner with EC50 values 9.47 μM and 10.48 μM, for USA-WA1/2020 and B.1.617.2 variants, respectively. Our results suggest that dasabuvir can be further evaluated as a therapeutic drug for COVID-19. Importantly, this system provides a robust, target-specific, and high-throughput screening compatible (z- and z’-factors of >0.5) platforms that will be a valuable tool for screening SARS-CoV-2 RdRp inhibitors.http://www.sciencedirect.com/science/article/pii/S2772892722000438SARS-CoV-2COVID-19RdRpAntiviralsDasabuvirRibavirin |
| spellingShingle | Timsy Uppal Kai Tuffo Svetlana Khaiboullina Sivani Reganti Mark Pandori Subhash C. Verma Screening of SARS-CoV-2 antivirals through a cell-based RNA-dependent RNA polymerase (RdRp) reporter assay Cell Insight SARS-CoV-2 COVID-19 RdRp Antivirals Dasabuvir Ribavirin |
| title | Screening of SARS-CoV-2 antivirals through a cell-based RNA-dependent RNA polymerase (RdRp) reporter assay |
| title_full | Screening of SARS-CoV-2 antivirals through a cell-based RNA-dependent RNA polymerase (RdRp) reporter assay |
| title_fullStr | Screening of SARS-CoV-2 antivirals through a cell-based RNA-dependent RNA polymerase (RdRp) reporter assay |
| title_full_unstemmed | Screening of SARS-CoV-2 antivirals through a cell-based RNA-dependent RNA polymerase (RdRp) reporter assay |
| title_short | Screening of SARS-CoV-2 antivirals through a cell-based RNA-dependent RNA polymerase (RdRp) reporter assay |
| title_sort | screening of sars cov 2 antivirals through a cell based rna dependent rna polymerase rdrp reporter assay |
| topic | SARS-CoV-2 COVID-19 RdRp Antivirals Dasabuvir Ribavirin |
| url | http://www.sciencedirect.com/science/article/pii/S2772892722000438 |
| work_keys_str_mv | AT timsyuppal screeningofsarscov2antiviralsthroughacellbasedrnadependentrnapolymeraserdrpreporterassay AT kaituffo screeningofsarscov2antiviralsthroughacellbasedrnadependentrnapolymeraserdrpreporterassay AT svetlanakhaiboullina screeningofsarscov2antiviralsthroughacellbasedrnadependentrnapolymeraserdrpreporterassay AT sivanireganti screeningofsarscov2antiviralsthroughacellbasedrnadependentrnapolymeraserdrpreporterassay AT markpandori screeningofsarscov2antiviralsthroughacellbasedrnadependentrnapolymeraserdrpreporterassay AT subhashcverma screeningofsarscov2antiviralsthroughacellbasedrnadependentrnapolymeraserdrpreporterassay |