@article{Rihn_S-2021_26422, title = {A plasmid DNA-launched SARS-CoV-2 reverse genetics system and coronavirus toolkit for COVID-19 research}, author = {Rihn, S. and Merits, A. and Bakshi, S. and Turnbull, M. and Wickenhagen, A. and Alexander, A. and Baillie, C. and Brennan, B. and Brown, F. and Brunker, K. and Bryden, S. and Burness, K. and Carmichael, S. and Cole, S. and Cowton, V. and Davies, P. and Davis, C. and De Lorenzo, G. and Donald, C. and Dorward, M. and Dunlop, J. and Elliott, M. and Fares, M. and da Silva Filipe, A. and Freitas, J. and Furnon, W. and Gestuveo, R. and Geyer, A. and Giesel, D. and Goldfarb, D. and Goodman, N. and Gunson, R. and Hastie, C. and Herder, V. and Hughes, J. and Johnson, C. and Johnson, N. and Kohl, A. and Kerr, K. and Leech, H. and Lello, L. and Li, K. and Lieber, G. and Liu, X. and Lingala, R. and Loney, C. and Mair, D. and McElwee, M. and McFarlane, S. and Nichols, J. and Nomikou, K. and Orr, A. and Orton, R. and Palmarini, M. and Parr, Y. and Pinto, R. and Raggett, S. and Reid, E. and Robertson, D. and Royle, J. and Cameron-Ruiz, N. and Shepherd, J. and Smollett, K. and Stewart, D. and Stewart, M. and Sugrue, E. and Szemiel, A. and Taggart, A. and Thomson, E. and Tong, L. and Torrie, L. and Toth, R. and Varjak, M. and Wang, S. and Wilkinson, S. and Wyatt, P. and Zusinaite, E. and Alessi, D. and Patel, A. and Zaid, A. and Wilson, S. and Mahalingam, S.}, month = {feb}, year = {2021}, 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.}, volume = {19}, issue = {2}, journal = {PLOS Biology}, publisher = {Public Library of Science}, url = {https://doi.org/10.1371/journal.pbio.3001091}, }