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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
general
Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform
Nature, Volume 582, No. 7813, Year 2020
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Description
Reverse genetics has been an indispensable tool to gain insights into viral pathogenesis and vaccine development. The genomes of large RNA viruses, such as those from coronaviruses, are cumbersome to clone and manipulate in Escherichia coli owing to the size and occasional instability of the genome1–3. Therefore, an alternative rapid and robust reverse-genetics platform for RNA viruses would benefit the research community. Here we show the full functionality of a yeast-based synthetic genomics platform to genetically reconstruct diverse RNA viruses, including members of the Coronaviridae, Flaviviridae and Pneumoviridae families. Viral subgenomic fragments were generated using viral isolates, cloned viral DNA, clinical samples or synthetic DNA, and these fragments were then reassembled in one step in Saccharomyces cerevisiae using transformation-associated recombination cloning to maintain the genome as a yeast artificial chromosome. T7 RNA polymerase was then used to generate infectious RNA to rescue viable virus. Using this platform, we were able to engineer and generate chemically synthesized clones of the virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)4, which has caused the recent pandemic of coronavirus disease (COVID-19), in only a week after receipt of the synthetic DNA fragments. The technical advance that we describe here facilitates rapid responses to emerging viruses as it enables the real-time generation and functional characterization of evolving RNA virus variants during an outbreak. © 2020, The Author(s), under exclusive licence to Springer Nature Limited.
Authors & Co-Authors
Labroussaa, Fabien
Switzerland, Bern
University of Bern
Portmann, Jasmine
Switzerland, Mittelhausen
Institut Für Viruskrankheiten Und Immunprophylaxe
Switzerland, Bern
University of Bern
Gultom, Mitra L.
Switzerland, Mittelhausen
Institut Für Viruskrankheiten Und Immunprophylaxe
Switzerland, Bern
University of Bern
Pfaender, Stephanie
Switzerland, Mittelhausen
Institut Für Viruskrankheiten Und Immunprophylaxe
Switzerland, Bern
University of Bern
Germany, Bochum
Ruhr-universitat Bochum
Cippà, Valentina L.
Switzerland, Bern
University of Bern
Muth, Doreen
Germany, Berlin
Freie Universität Berlin
Germany, Berlin
Charité – Universitätsmedizin Berlin
Niemeyer, Daniela
Germany, Berlin
Freie Universität Berlin
Germany, Berlin
Charité – Universitätsmedizin Berlin
Corman, Victor Max
Germany, Berlin
Freie Universität Berlin
Germany, Berlin
Charité – Universitätsmedizin Berlin
Müller, Marcel A.
Germany, Berlin
Freie Universität Berlin
Germany, Berlin
Charité – Universitätsmedizin Berlin
Russian Federation, Moscow
Sechenov First Moscow State Medical University
Drosten, C.
Germany, Berlin
Freie Universität Berlin
Germany, Berlin
Charité – Universitätsmedizin Berlin
Dijkman, Ronald
Switzerland, Mittelhausen
Institut Für Viruskrankheiten Und Immunprophylaxe
Switzerland, Bern
University of Bern
Jores, Joerg
Switzerland, Bern
University of Bern
Thiel, Volker
Switzerland, Mittelhausen
Institut Für Viruskrankheiten Und Immunprophylaxe
Switzerland, Bern
University of Bern
Statistics
Citations: 269
Authors: 13
Affiliations: 6
Identifiers
Doi:
10.1038/s41586-020-2294-9
ISSN:
00280836
Research Areas
Cancer
Covid
Genetics And Genomics