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

Tran, Thi Nhu Thao; Labroussaa, Fabien; Ebert, Nadine; V'kovski, Philip; Stalder, Hanspeter; Portmann, Jasmine; Kelly, Jenna; Steiner, Silvio; Holwerda, Melle; Kratzel, Annika; Gultom, Mitra; Schmied, Kimberly; Laloli, Laura; Hüsser, Linda; Wider, Manon; Pfaender, Stephanie; Hirt, Dagny; Cippà, Valentina; Crespo-Pomar, Silvia; Schröder, Simon; ... (2020). Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform. Nature, 582(7813), pp. 561-565. Springer Nature 10.1038/s41586-020-2294-9

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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.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Research
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Tran, Thi Nhu Thao; Labroussaa, Fabien; Ebert, Nadine; V'kovski, Philip; Stalder, Hanspeter; Portmann, Jasmine; Kelly, Jenna Nicole; Steiner, Silvio; Holwerda, Melle; Kratzel, Annika; Gultom, Mitra Lovelin; Schmied, Kimberly Shadia; Laloli, Laura; Hüsser, Linda; Wider, Manon Flore; Pfänder, Stephanie; Hirt, Dagny Nora; Cippà, Valentina Lucia; Crespo Pomar, Silvia; Dijkman, Ronald; Jores, Jörg and Thiel, Volker Earl

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health
600 Technology > 630 Agriculture

ISSN:

1476-4687

Publisher:

Springer Nature

Language:

English

Submitter:

Achim Braun Parham

Date Deposited:

30 Jul 2020 09:23

Last Modified:

30 Jul 2020 09:23

Publisher DOI:

10.1038/s41586-020-2294-9

PubMed ID:

32365353

BORIS DOI:

10.7892/boris.145427

URI:

https://boris.unibe.ch/id/eprint/145427

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