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Abstract
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) virus, which is highly pathogenic and classified as a biosafety level 3 (BSL-3) agent, has greatly threatened global health and efficacious antivirals are urgently needed. The high requirement of facilities to manipulate the live virus has limited the development of antiviral study. Here, we constructed a reporter replicon of SARS-CoV-2, which can be handled in a BSL-2 laboratory. The Renilla luciferase activity effectively reflected the transcription and replication levels of the replicon genome. We identified the suitability of the replicon in antiviral screening using the known inhibitors, and thus established the replicon-based high-throughput screening (HTS) assay for SARS-CoV-2. The application of the HTS assay was further validated using a few hit natural compounds, which were screened out in a SARS-CoV-2 induced cytopathic-effect-based HTS assay in our previous study. This replicon-based HTS assay will be a safe platform for SARS-CoV-2 antiviral screening in a BSL-2 laboratory without the live virus.
- Received:
- Accepted:
- Published Online:
Funding
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the Education Department Program of Hunan Province, China
(Award 17C0962)
- Principle Award Recipient: Xiao-DanLi
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Guangzhou Institute of Pediatrics, Guangzhou Women and Children’s Medical Center
(Award IP-2019-002)
- Principle Award Recipient: YiXu
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Guangzhou Institute of Pediatrics, Guangzhou Women and Children’s Medical Center
(Award Pre-NSFC-2018-007)
- Principle Award Recipient: YiXu
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Wuhan Science and technology project
(Award 2018201261638501)
- Principle Award Recipient: BoZhang
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the National major science and technology project
(Award 2018ZX10711001-006)
- Principle Award Recipient: BoZhang