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. Author manuscript; available in PMC: 2021 Sep 1.
Published in final edited form as: Nat Protoc. 2021 Jan 29;16(3):1761–1784. doi: 10.1038/s41596-021-00491-8

Figure 1. Overview of SARS-CoV-2 Reverse Genetic Systems.

Figure 1.

The SARS-CoV-2 infectious clone model contains seven cDNA fragments to cover the complete viral genome, to disrupt toxic elements, and to aid in genetic manipulation. The SARS-CoV-2 plasmids are amplified in E. coli and sequentially ligated following digestion with type II restriction enzymes to remove the plasmid backbone. The full-length viral DNA is then in vitro transcribed using T7 polymerase to generate full-length genomic SARS-CoV-2 RNA and electroporated into cells with N-protein transcript expressed in trans. Following electroporation, cells are seeded into cell culture flasks and virus recovered 2–5 days post electroporation.