A plasmid DNA-launched SARS-CoV-2 reverse genetics system and coronavirus toolkit for COVID-19 research

Rihn, Suzannah J. and Merits, Andres and Bakshi, Siddharth and Turnbull, Matthew L. and Wickenhagen, Arthur and Alexander, Akira J. T. and Baillie, Carla and Brennan, Benjamin and Brown, Fiona and Brunker, Kirstyn and Bryden, Steven R. and Burness, Kerry A. and Carmichael, Stephen and Cole, Sarah J. and Cowton, Vanessa M. and Davies, Paul and Davis, Chris and De Lorenzo, Giuditta and Donald, Claire L. and Dorward, Mark and Dunlop, James I. and Elliott, Matthew and Fares, Mazigh and da Silva Filipe, Ana and Freitas, Joseph R. and Furnon, Wilhelm and Gestuveo, Rommel J. and Geyer, Anna and Giesel, Daniel and Goldfarb, Daniel M. and Goodman, Nicola and Gunson, Rory and Hastie, C. James and Herder, Vanessa and Hughes, Joseph and Johnson, Clare and Johnson, Natasha and Kohl, Alain and Kerr, Karen and Leech, Hannah and Lello, Laura Sandra and Li, Kathy and Lieber, Gauthier and Liu, Xiang and Lingala, Rajendra and Loney, Colin and Mair, Daniel and McElwee, Marion J. and McFarlane, Steven and Nichols, Jenna and Nomikou, Kyriaki and Orr, Anne and Orton, Richard J. and Palmarini, Massimo and Parr, Yasmin A. and Pinto, Rute Maria and Raggett, Samantha and Reid, Elaine and Robertson, David L. and Royle, Jamie and Cameron-Ruiz, Natalia and Shepherd, James G. and Smollett, Katherine and Stewart, Douglas G. and Stewart, Meredith and Sugrue, Elena and Szemiel, Agnieszka M. and Taggart, Aislynn and Thomson, Emma C. and Tong, Lily and Torrie, Leah S. and Toth, Rachel and Varjak, Margus and Wang, Sainan and Wilkinson, Stuart G. and Wyatt, Paul G. and Zusinaite, Eva and Alessi, Dario R. and Patel, Arvind H. and Zaid, Ali and Wilson, Sam J. and Mahalingam, Suresh and Cimarelli, Andrea (2021) A plasmid DNA-launched SARS-CoV-2 reverse genetics system and coronavirus toolkit for COVID-19 research. PLOS Biology, 19 (2). e3001091. ISSN 1545-7885

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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.

Item Type: Article
Subjects: Oalibrary Press > Biological Science
Depositing User: Managing Editor
Date Deposited: 14 Mar 2023 09:04
Last Modified: 06 May 2024 06:16
URI: http://asian.go4publish.com/id/eprint/795

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