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

PLoS Biol . 2021-02; 
Suzannah J Rihn, Andres Merits, Siddharth Bakshi, Matthew L Turnbull, Arthur Wickenhagen, Akira J T Alexander,
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Gene Synthesis (A) Schematic of the design and construction of a pCC1-4K-SARS-CoV2-Wuhan-Hu-1 icDNA clone. Synthetic DNA fragments 1, 2, and 3 based on the SARS-CoV-2-Wuhan-Hu1 sequence were cloned into the pCC1 plasmids, and fragments 4 and 5 were cloned into high copy plasmid pUC57Kan by the gene synthesis company (Genscript). Get A Quote

摘要

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 ... More

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