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

Author

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

Subjects
Biology (General), QH301-705.5
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.

Published
2021
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2. A plasmid DNA-launched SARS-CoV-2 reverse genetics system and coronavirus toolkit for COVID-19 research

Author

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

Subjects
0301 basic medicine, RNA viruses, Viral Diseases, Pulmonology, Coronaviruses, Physiology, viruses, medicine.disease_cause, Biochemistry, Mice, 0302 clinical medicine, Plasmid, Medical Conditions, Immune Physiology, Pandemic, Chlorocebus aethiops, Biology (General), skin and connective tissue diseases, Pathology and laboratory medicine, Coronavirus, Immune System Proteins, General Neuroscience, Serine Endopeptidases, Methods and Resources, virus diseases, Medical microbiology, 3. Good health, Precipitation Techniques, Infectious Diseases, Viruses, Angiotensin-Converting Enzyme 2, Antibody, SARS CoV 2, Pathogens, General Agricultural and Biological Sciences, Plasmids, COVID-19 Vaccines, SARS coronavirus, Viral protein, QH301-705.5, Morpholines, Immunology, Biology, Research and Analysis Methods, Microbiology, General Biochemistry, Genetics and Molecular Biology, Antibodies, 03 medical and health sciences, Respiratory Disorders, Open Reading Frames, Viral Proteins, medicine, Immunoprecipitation, Animals, Humans, Codon, Vero Cells, Medicine and health sciences, SARS, General Immunology and Microbiology, Biology and life sciences, SARS-CoV-2, fungi, Organisms, Viral pathogens, Hydrazones, Proteins, COVID-19, Covid 19, Virology, Reverse genetics, Reverse Genetics, Microbial pathogens, body regions, Open reading frame, 030104 developmental biology, Pyrimidines, A549 Cells, Respiratory Infections, Vero cell, biology.protein, 030217 neurology & neurosurgery
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., To help meet the ensuing demand for COVID-19 reagents, this article presents an openly available ‘coronavirus toolkit’ (https://mrcppu-covid.bio) and describes the generation and validation of these tools, including a simple SARS-CoV-2 reverse genetics system and a near-comprehensive panel of SARS-CoV-2 antibodies.

Published
2021

3. Visitors' Book The Singing Tower Mountain Lake

Author

Bok, Curtis, 1897-1962; Bok, Cary William; Bok, Margaret Plummer; Bok, Nellie Lee Holt, 1901-1984; Coolidge, Calvin, 1872-1933; Coolidge, Grace Goodhue, 1879-1957; Carlton, Doyle Elam, 1885-1972; Carlton, Nell Ray, 1891-1982; Sanders, Everett, 1882-1950; Williams, Harry H.; Williams, Marim P.; Taylor, E. Denison; Medary, Milton Bennett, 1874-1929; Medary, Hannah Leech Stadelman, 1875-; Yellin, Samuel, 1885-1940; Burrell, Horace H. (1894-1936); Brees, Anton; Nornabell, Harry M.; Nornabell, Catherine SEE: Van Dyke, Catherine; Nornabell, Ursula; Van Dyke, Catherine; Olmsted, Frederick Law, 1870-1957, Bok, Edward William, 1863-1930; Bok, Mary Louise SEE: Zimbalist, Mary Louise Curtis Bok, 1876-1970; Zimbalist, Mary Louise Curtis Bok, 1876-1970, Bok, Curtis, 1897-1962; Bok, Cary William; Bok, Margaret Plummer; Bok, Nellie Lee Holt, 1901-1984; Coolidge, Calvin, 1872-1933; Coolidge, Grace Goodhue, 1879-1957; Carlton, Doyle Elam, 1885-1972; Carlton, Nell Ray, 1891-1982; Sanders, Everett, 1882-1950; Williams, Harry H.; Williams, Marim P.; Taylor, E. Denison; Medary, Milton Bennett, 1874-1929; Medary, Hannah Leech Stadelman, 1875-; Yellin, Samuel, 1885-1940; Burrell, Horace H. (1894-1936); Brees, Anton; Nornabell, Harry M.; Nornabell, Catherine SEE: Van Dyke, Catherine; Nornabell, Ursula; Van Dyke, Catherine; Olmsted, Frederick Law, 1870-1957, and Bok, Edward William, 1863-1930; Bok, Mary Louise SEE: Zimbalist, Mary Louise Curtis Bok, 1876-1970; Zimbalist, Mary Louise Curtis Bok, 1876-1970

Abstract

Day of Dedication of the Sanctuary and the Singing Tower by the President

Published
2018

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