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Construction of a recombinant porcine epidemic diarrhea virus encoding nanoluciferase for high-throughput screening of natural antiviral products
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Document Title
Construction of a recombinant porcine epidemic diarrhea virus encoding nanoluciferase for high-throughput screening of natural antiviral products
Author
Li W.,Zhang M.,Zheng H.,Zhou P.,Liu Z.,Jongkaewwattana A.,Luo R.,He Q.
Name from Authors Collection
Affiliations
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, the Cooperative Innovation Center for Sustainable Pig Production, Wuhan, 430070, China; Key Laboratory of Pesticide & Chemical Biology of Ministry of Education, Hubei International Scientific and Technological Cooperation Base of Pesticide and Green Synthesis, International Joint Research Center for Intelligent Biosensing Technology and Health, College of Chemistry, Central China Normal University, Wuhan, 430079, China; Virology and Cell Technology Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Phahonyothin Rd., Pathumthani, 12120, Thailand
Type
Article
Source Title
Viruses
ISSN
19994915
Year
2021
Volume
13
Issue
9
Open Access
All Open Access, Gold, Green
Publisher
MDPI
DOI
10.3390/v13091866
Abstract
Porcine epidemic diarrhea virus (PEDV) is the predominant cause of an acute, highly con-tagious enteric disease in neonatal piglets. There are currently no approved drugs against PEDV infection. Here, we report the development of a nanoluciferase (NLuc)-based high-throughput screening (HTS) platform to identify novel anti-PEDV compounds. We constructed a full-length cDNA clone for a cell-adapted PEDV strain YN150. Using reverse genetics, we replaced the open reading frame 3 (ORF3) in the viral genome with an NLuc gene to engineer a recombinant PEDV expressing NLuc (rPEDV-NLuc). rPEDV-NLuc produced similar plaque morphology and showed similar growth kinetics compared with the wild-type PEDV in vitro. Remarkably, the level of lucif-erase activity could be stably detected in rPEDV-NLuc-infected cells and exhibited a strong positive correlation with the viral titers. Given that NLuc expression represents a direct readout of PEDV replication, anti-PEDV compounds could be easily identified by quantifying the NLuc activity. Using this platform, we screened for the anti-PEDV compounds from a library of 803 natural products and identified 25 compounds that could significantly inhibit PEDV replication. Interestingly, 7 of the 25 identified compounds were natural antioxidants, including Betulonic acid, Ursonic acid, es-culetin, lithocholic acid, nordihydroguaiaretic acid, caffeic acid phenethyl ester, and grape seed ex-tract. As expected, all of the antioxidants could potently reduce PEDV-induced oxygen species pro-duction, which, in turn, inhibit PEDV replication in a dose-dependent manner. Collectively, our findings provide a powerful platform for the rapid screening of promising therapeutic compounds against PEDV infection. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
Funding Sponsor
National Natural Science Foundation of China
License
CC BY
Rights
Author
Publication Source
Scopus