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A plasmid DNA-launched SARS-CoV-2 reverse genetics system and coronavirus toolkit for COVID-19 research.
Rihn, Suzannah J; Merits, Andres; Bakshi, Siddharth; Turnbull, Matthew L; Wickenhagen, Arthur; Alexander, Akira J T; Baillie, Carla; Brennan, Benjamin; Brown, Fiona; Brunker, Kirstyn; Bryden, Steven R; Burness, Kerry A; Carmichael, Stephen; Cole, Sarah J; Cowton, Vanessa M; Davies, Paul; Davis, Chris; De Lorenzo, Giuditta; Donald, Claire L; Dorward, Mark; Dunlop, James I; Elliott, Matthew; Fares, Mazigh; da Silva Filipe, Ana; Freitas, Joseph R; Furnon, Wilhelm; Gestuveo, Rommel J; Geyer, Anna; Giesel, Daniel; Goldfarb, Daniel M; Goodman, Nicola; Gunson, Rory; Hastie, C James; Herder, Vanessa; Hughes, Joseph; Johnson, Clare; Johnson, Natasha; Kohl, Alain; Kerr, Karen; Leech, Hannah; Lello, Laura Sandra; Li, Kathy; Lieber, Gauthier; Liu, Xiang; Lingala, Rajendra; Loney, Colin; Mair, Daniel; McElwee, Marion J; McFarlane, Steven; Nichols, Jenna.
  • Rihn SJ; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Merits A; Institute of Technology, University of Tartu, Tartu, Estonia.
  • Bakshi S; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Turnbull ML; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Wickenhagen A; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Alexander AJT; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Baillie C; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Brennan B; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Brown F; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Brunker K; Institute of Biodiversity, Animal Health and Comparative Medicine, University of Glasgow, Glasgow, United Kingdom.
  • Bryden SR; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Burness KA; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Carmichael S; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Cole SJ; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Cowton VM; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Davies P; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Davis C; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • De Lorenzo G; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Donald CL; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Dorward M; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Dunlop JI; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Elliott M; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Fares M; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • da Silva Filipe A; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Freitas JR; Emerging Viruses, Inflammation and Therapeutics Group, Menzies Health Institute Queensland, Griffith University, Gold Coast, Queensland, Australia.
  • Furnon W; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Gestuveo RJ; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Geyer A; Division of Biological Sciences, College of Arts and Sciences, University of the Philippines Visayas, Miagao, Iloilo, Philippines.
  • Giesel D; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Goldfarb DM; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Goodman N; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Gunson R; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Hastie CJ; West of Scotland Specialist Virology Centre, Glasgow, United Kingdom.
  • Herder V; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Hughes J; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Johnson C; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Johnson N; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Kohl A; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Kerr K; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Leech H; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Lello LS; MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • Li K; Institute of Technology, University of Tartu, Tartu, Estonia.
  • Lieber G; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Liu X; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Lingala R; Emerging Viruses, Inflammation and Therapeutics Group, Menzies Health Institute Queensland, Griffith University, Gold Coast, Queensland, Australia.
  • Loney C; Indian Immunologicals Ltd (IIL), Rakshapuram, Gachibowli Post, Hyderabad Telangana, India.
  • Mair D; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • McElwee MJ; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • McFarlane S; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
  • Nichols J; MRC-University of Glasgow Centre for Virus Research (CVR), Glasgow, United Kingdom.
PLoS Biol ; 19(2): e3001091, 2021 02.
Article in English | MEDLINE | ID: covidwho-1102372
Semantic information from SemMedBD (by NLM)
1. Antibodies PART_OF 2019 novel coronavirus
Subject
Antibodies
Predicate
PART_OF
Object
2019 novel coronavirus
2. Apilimod STIMULATES TMPRSS2 gene|TMPRSS2
Subject
Apilimod
Predicate
STIMULATES
Object
TMPRSS2 gene|TMPRSS2
3. Antibodies PART_OF 2019 novel coronavirus
Subject
Antibodies
Predicate
PART_OF
Object
2019 novel coronavirus
4. Apilimod STIMULATES TMPRSS2 gene|TMPRSS2
Subject
Apilimod
Predicate
STIMULATES
Object
TMPRSS2 gene|TMPRSS2
ABSTRACT
The recent emergence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the underlying cause of Coronavirus Disease 2019 (COVID-19), has led to a worldwide pandemic causing substantial morbidity, mortality, and economic devastation. In response, many laboratories have redirected attention to SARS-CoV-2, meaning there is an urgent need for tools that can be used in laboratories unaccustomed to working with coronaviruses. Here we report a range of tools for SARS-CoV-2 research. First, we describe a facile single plasmid SARS-CoV-2 reverse genetics system that is simple to genetically manipulate and can be used to rescue infectious virus through transient transfection (without in vitro transcription or additional expression plasmids). The rescue system is accompanied by our panel of SARS-CoV-2 antibodies (against nearly every viral protein), SARS-CoV-2 clinical isolates, and SARS-CoV-2 permissive cell lines, which are all openly available to the scientific community. Using these tools, we demonstrate here that the controversial ORF10 protein is expressed in infected cells. Furthermore, we show that the promising repurposed antiviral activity of apilimod is dependent on TMPRSS2 expression. Altogether, our SARS-CoV-2 toolkit, which can be directly accessed via our website at https//mrcppu-covid.bio/, constitutes a resource with considerable potential to advance COVID-19 vaccine design, drug testing, and discovery science.
Subject(s)

Full text: Available Collection: International databases Database: MEDLINE Main subject: Reverse Genetics / COVID-19 Vaccines / SARS-CoV-2 / COVID-19 Topics: Vaccines Limits: Animals / Humans Language: English Journal: PLoS Biol Journal subject: Biology Year: 2021 Document Type: Article Affiliation country: Journal.pbio.3001091

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Reverse Genetics / COVID-19 Vaccines / SARS-CoV-2 / COVID-19 Topics: Vaccines Limits: Animals / Humans Language: English Journal: PLoS Biol Journal subject: Biology Year: 2021 Document Type: Article Affiliation country: Journal.pbio.3001091