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

Nature. 2020-05; 
Tran Thi Nhu Thao, Fabien Labroussaa, Nadine Ebert, Philip V’kovski, Hanspeter Stalder, Jasmine Portmann, Jenna Kelly, Silvio Steiner, Melle Holwerda, Annika Kratzel, Mitra Gultom, Laura Laloli, Linda Hüsser, Manon Wider, Stephanie Pfaender, Dagny Hirt, Valentina Cippà, Silvia Crespo-Pomar, Simon Schröder, Doreen Muth, Daniela Niemeyer, Marcel A. Müller, Christian Drosten, Ronald Dijkman, Joerg Jores, Volker Thiel
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Gene Synthesis We thank the J. Craig Venter Institute (JCVI) for provision of the TAR vectors. We thank specifically Eric Sun and Tibbers Zhang and are grateful for the timely provision of the synthetic DNA fragments from GenScript. Get A Quote

Abstract

Reverse genetics has been an indispensable tool revolutionising our insights into viral pathogenesis and vaccine development. Large RNA virus genomes, such as from Coronavirus, are cumbersome to clone and to manipulate in E. coli hosts due to size and occasional instability1-3. 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 for the genetic reconstruction of diverse RNA viruses, including members of the Coronaviridae, Flaviviridae and Paramyxoviridae families. Viral subgenomic fragments were generated using viral isolates, cloned viral DNA, clinical sa... More

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