TY - JOUR
AU - Rihn, Suzannah J.
AU - Merits, Andreas
AU - Bakshi, Siddharth
AU - Turnbull, Matthew L.
AU - Wickenhagen, Arthur
AU - Alexander, Akira J.T.
AU - Baillie, Carla
AU - Brennan, Benjamin
AU - Brown, Fiona
AU - Brunker, Kirstyn
AU - Bryden, Steven R.
AU - Burness, Kerry A.
AU - Carmichael, Stephen
AU - Cole, Sarah J.
AU - Cowton, Vanessa M.
AU - Davies, Paul
AU - Davis, Christopher
AU - de Lorenzo, Giuditta
AU - Donald, Claire L.
AU - Dorward, Mark
AU - Dunlop, James I.
AU - Elliott, Matthew
AU - Fares, Mazigh
AU - da Silva Filipe, Ana
AU - Freitas, Joseph R.
AU - Furnon, Wilhelm
AU - Gestuveo, Rommel J.
AU - Geyer, Anna
AU - Giesel, Daniel
AU - Goldfarb, Daniel M.
AU - Goodman, Nicola
AU - Gunson, Rory N.
AU - Hastie, C. James
AU - Herder, Vanessa
AU - Hughes, Joseph
AU - Johnson, Clare
AU - Johnson, Natasha
AU - Kohl, Alain
AU - Kerr, Karen
AU - Leech, Hannah
AU - Lello, Laura Sandra
AU - Li, Kathy K.
AU - Lieber, Gauthier
AU - Liu, Xiang
AU - Lingala, Rajendra
AU - Loney, Colin
AU - Mair, Daniel
AU - McElwee, Marion J.
AU - McFarlane, Steven
AU - Nichols, Jenna
AU - Nomikou, Kyriaki
AU - Orr, Anne
AU - Orton, Richard J.
AU - Palmarini, Massimo
AU - Parr, Yasmin A.
AU - Pinto, Rute Maria
AU - Raggett, Samantha
AU - Reid, Elaine
AU - Robertson, David L.
AU - Royle, Jamie
AU - Cameron-Ruiz, Natalia
AU - Shepherd, James G.
AU - Smollett, Katherine
AU - Stewart, Douglas G.
AU - Stewart, Meredith
AU - Sugrue, Elena
AU - Szemiel, Agnieszka
AU - Taggart, Aislynn
AU - Thomson, Emma C.
AU - Tong, Lily
AU - Torrie, Leah S.
AU - Toth, Rachel
AU - Varjak, Margus
AU - Wang, Sainan
AU - Wilkinson, Stuart G.
AU - Wyatt, Paul Graham
AU - Zusinaite, Eva
AU - Alessi, Dario R.
AU - Patel, Arvind H.
AU - Zaid, Ali
AU - Wilson, Sam J.
AU - Mahalingam, Suresh
PY - 2021
DA - February
TI - A plasmid DNA-launched SARS-CoV-2 reverse genetics system and coronavirus toolkit for COVID-19 research
JO - PLOS Biology
VL - 19
IS - 2
DO - https://doi.org/10.1371/journal.pbio.3001091
AB - 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.
PB - Public Library of Science
UR - http://eprints.gla.ac.uk/232858/
KW - Coronavirus (COVID-19)
ER