Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform

Tran Thao, Fabien Labroussaa, Nadine Ebert, Philip V’kovski, Hanspeter Stalder, Jasmine Portmann, Jenna Kelly, Silvio Steiner, Melle Holwerda, Annika Kratzel, Mitra Gultom, Kimberly Schmied, Laura Laloli, Linda Hüsser, Manon Wider, Stephanie Pfaender, Dagny Hirt, Valentina Cippà, Silvia Crespo-Pomar, Simon Schröder, Doreen Muth, Daniela Niemeyer, Victor M. Corman, Marcel A. Müller, Christian Drosten, Ronald Dijkman, Joerg Jores () and Volker Thiel ()
Additional contact information
Tran Thao: Institute of Virology and Immunology (IVI)
Fabien Labroussaa: University of Bern
Nadine Ebert: Institute of Virology and Immunology (IVI)
Philip V’kovski: Institute of Virology and Immunology (IVI)
Hanspeter Stalder: Institute of Virology and Immunology (IVI)
Jasmine Portmann: Institute of Virology and Immunology (IVI)
Jenna Kelly: Institute of Virology and Immunology (IVI)
Silvio Steiner: Institute of Virology and Immunology (IVI)
Melle Holwerda: Institute of Virology and Immunology (IVI)
Annika Kratzel: Institute of Virology and Immunology (IVI)
Mitra Gultom: Institute of Virology and Immunology (IVI)
Kimberly Schmied: Institute of Virology and Immunology (IVI)
Laura Laloli: Institute of Virology and Immunology (IVI)
Linda Hüsser: Institute of Virology and Immunology (IVI)
Manon Wider: University of Bern
Stephanie Pfaender: Institute of Virology and Immunology (IVI)
Dagny Hirt: Institute of Virology and Immunology (IVI)
Valentina Cippà: University of Bern
Silvia Crespo-Pomar: University of Bern
Simon Schröder: corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health
Doreen Muth: corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health
Daniela Niemeyer: corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health
Victor M. Corman: corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health
Marcel A. Müller: corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health
Christian Drosten: corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health
Ronald Dijkman: Institute of Virology and Immunology (IVI)
Joerg Jores: University of Bern
Volker Thiel: Institute of Virology and Immunology (IVI)

Nature, 2020, vol. 582, issue 7813, 561-565

Abstract: Abstract 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.

Date: 2020
References: Add references at CitEc
Citations: View citations in EconPapers (9)

Downloads: (external link)
https://www.nature.com/articles/s41586-020-2294-9 Abstract (text/html)
Access to the full text of the articles in this series is restricted.

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:nature:v:582:y:2020:i:7813:d:10.1038_s41586-020-2294-9

Ordering information: This journal article can be ordered from
https://www.nature.com/

DOI: 10.1038/s41586-020-2294-9

Access Statistics for this article

Nature is currently edited by Magdalena Skipper

More articles in Nature from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().