Construction of a Noninfectious SARS-CoV-2 Replicon for Antiviral-Drug Testing and Gene Function Studies.
SARS-CoV-2
antivirals
nano luciferase
nonstructural protein 15
nucleocapsid protein
replicon
reverse genetics
Journal
Journal of virology
ISSN: 1098-5514
Titre abrégé: J Virol
Pays: United States
ID NLM: 0113724
Informations de publication
Date de publication:
25 08 2021
25 08 2021
Historique:
pubmed:
1
7
2021
medline:
7
9
2021
entrez:
30
6
2021
Statut:
ppublish
Résumé
The emerging coronavirus disease 2019 (COVID-19) outbreak caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly spread worldwide, resulting in global public health emergencies and economic crises. In the present study, a noninfectious and biosafety level 2 (BSL2)-compatible SARS-CoV-2 replicon expressing a nano luciferase (nLuc) reporter was constructed in a bacterial artificial chromosomal (BAC) vector by reverse genetics. The nLuc reporter is highly sensitive, easily quantifiable, and high throughput adaptable. Upon transfecting the SARS-CoV-2 replicon BAC plasmid DNA into Vero E6 cells, we could detect high levels of nLuc reporter activity and viral RNA transcript, suggesting the replication of the replicon. The replicon replication was further demonstrated by the findings that deleting nonstructural protein 15 or mutating its catalytic sites significantly reduced replicon replication, whereas providing the nucleocapsid protein in
Identifiants
pubmed: 34191580
doi: 10.1128/JVI.00687-21
pmc: PMC8387049
doi:
Substances chimiques
Antiviral Agents
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0068721Subventions
Organisme : NIDCD NIH HHS
ID : F32 DC000190
Pays : United States
Organisme : Gouvernement du Canada | Canadian Institutes of Health Research (CIHR)
ID : DC0190GP
Organisme : Saskatchewan Agriculture Development Fund
ID : 20180101
Organisme : Gouvernement du Canada | Natural Sciences and Engineering Research Council of Canada (NSERC)
ID : RGPIN-2018-04138
Références
Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8794-7
pubmed: 1528894
Proc Natl Acad Sci U S A. 2004 Aug 24;101(34):12694-9
pubmed: 15304651
Proc Natl Acad Sci U S A. 2017 May 23;114(21):E4251-E4260
pubmed: 28484023
Proc Natl Acad Sci U S A. 2000 May 9;97(10):5516-21
pubmed: 10805807
Viruses. 2014 Aug 07;6(8):2991-3018
pubmed: 25105276
Virology. 2007 Mar 30;360(1):150-8
pubmed: 17098272
J Gen Virol. 2005 Jan;86(Pt 1):211-215
pubmed: 15604448
J Virol. 2004 Nov;78(22):12683-8
pubmed: 15507657
mBio. 2020 Sep 25;11(5):
pubmed: 32978313
Methods Mol Biol. 2015;1282:135-52
pubmed: 25720478
Nature. 2020 Jul;583(7816):459-468
pubmed: 32353859
Vaccines (Basel). 2019 Apr 24;7(2):
pubmed: 31022829
Protein Sci. 2020 Jul;29(7):1596-1605
pubmed: 32304108
Science. 2020 Jun 19;368(6497):1331-1335
pubmed: 32321856
J Hum Genet. 2020 Dec;65(12):1075-1082
pubmed: 32699345
Virus Res. 2014 Aug 30;189:262-70
pubmed: 24930446
Front Neurol. 2020 Nov 16;11:583459
pubmed: 33304309
Biochem Biophys Res Commun. 2008 Sep 12;374(1):138-42
pubmed: 18619943
Gene Rep. 2020 Jun;19:100682
pubmed: 32300673
Sci Transl Med. 2017 Jun 28;9(396):
pubmed: 28659436
Antiviral Res. 2019 Sep;169:104541
pubmed: 31233808
Virology. 2018 Apr;517:157-163
pubmed: 29307596
Pathogens. 2020 Mar 04;9(3):
pubmed: 32143502
Inform Med Unlocked. 2020;20:100392
pubmed: 32835078
Proc Natl Acad Sci U S A. 2020 Mar 24;117(12):6771-6776
pubmed: 32054787
J Virol. 2020 Dec 23;:
pubmed: 33361429
N Engl J Med. 2020 Dec 31;383(27):2598-2600
pubmed: 33264539
Cell Host Microbe. 2020 May 13;27(5):841-848.e3
pubmed: 32289263
Cell Rep. 2020 Oct 6;33(1):108234
pubmed: 32979938
Int J Pept Res Ther. 2020 Nov 10;:1-16
pubmed: 33192207
Cell Host Microbe. 2020 Apr 8;27(4):671-680.e2
pubmed: 32183941
J Virol. 2007 Dec;81(24):13587-97
pubmed: 17898055
J Biomol Struct Dyn. 2020 Sep 4;:1-11
pubmed: 32885740
J Biomol Struct Dyn. 2021 Apr;39(7):2607-2616
pubmed: 32238094
Front Immunol. 2020 Oct 28;11:587615
pubmed: 33193414
NPJ Vaccines. 2020 Sep 28;5:91
pubmed: 33083026
Biochem Biophys Res Commun. 2014 Oct 3;452(4):1046-53
pubmed: 25242740
Proc Natl Acad Sci U S A. 2020 Apr 7;117(14):8094-8103
pubmed: 32198201
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
J Virol. 2020 Nov 9;94(23):
pubmed: 32938769
J Virol. 2015 Dec 09;90(4):2090-101
pubmed: 26656704
Phytomedicine. 2021 May;85:153317
pubmed: 32943302
Nature. 2020 Jun;582(7813):561-565
pubmed: 32365353
PLoS Biol. 2021 Feb 25;19(2):e3001091
pubmed: 33630831
Appl Soft Comput. 2020 Nov;96:106626
pubmed: 32834800
J Virol Methods. 2020 Oct;284:113926
pubmed: 32650037
Cell Rep. 2020 Jul 21;32(3):107940
pubmed: 32668216
J Virol. 2010 Feb;84(4):2169-75
pubmed: 19955314
Proc Natl Acad Sci U S A. 2014 Sep 16;111(37):E3900-9
pubmed: 25197083
J Virol. 2006 Nov;80(21):10900-6
pubmed: 16928748
Sci Rep. 2020 Aug 4;10(1):13093
pubmed: 32753646
Adv Exp Med Biol. 2006;581:579-83
pubmed: 17037601
PLoS Pathog. 2017 Feb 3;13(2):e1006195
pubmed: 28158275
PLoS Pathog. 2021 Feb 26;17(2):e1008690
pubmed: 33635931
Virol Sin. 2021 Apr 9;:
pubmed: 33835389
Biomed Pharmacother. 2020 Dec;132:110914
pubmed: 33254432