tetano
Editor, Senior Moderator
Cell Host Microbe. 2020 Apr 9. pii: S1931-3128(20)30231-6. doi: 10.1016/j.chom.2020.04.004. [Epub ahead of print]
An Infectious cDNA Clone of SARS-CoV-2.
Xie X[SUP]1[/SUP], Muruato A[SUP]2[/SUP], Lokugamage KG[SUP]3[/SUP], Narayanan K[SUP]3[/SUP], Zhang X[SUP]4[/SUP], Zou J[SUP]4[/SUP], Liu J[SUP]3[/SUP], Schindewolf C[SUP]3[/SUP], Bopp NE[SUP]5[/SUP], Aguilar PV[SUP]6[/SUP], Plante KS[SUP]7[/SUP], Weaver SC[SUP]8[/SUP], Makino S[SUP]9[/SUP], LeDuc JW[SUP]10[/SUP], Menachery VD[SUP]11[/SUP], Shi PY[SUP]12[/SUP].
Author information
Abstract
The ongoing pandemic of COVID-19, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), underscores the urgency to develop experimental systems for studying this virus and identifying countermeasures. We report a reverse genetic system for SARS-CoV-2. Seven complimentary DNA (cDNA) fragments spanning the SARS-CoV-2 genome were assembled into a full-genome cDNA. RNA transcribed from the full-genome cDNA was highly infectious after electroporation into cells, producing 2.9 ? 10[SUP]6[/SUP] plaque-forming unit (PFU)/mL of virus. Compared with a clinical isolate, the infectious-clone-derived SARS-CoV-2 (icSARS-CoV-2) exhibited similar plaque morphology, viral RNA profile, and replication kinetics. Additionally, icSARS-CoV-2 retained engineered molecular markers and did not acquire other mutations. We generated a stable mNeonGreen SARS-CoV-2 (icSARS-CoV-2-mNG) by introducing this reporter gene into ORF7 of the viral genome. icSARS-CoV-2-mNG was successfully used to evaluate the antiviral activities of interferon (IFN). Collectively, the reverse genetic system and reporter virus provide key reagents to study SARS-CoV-2 and develop countermeasures.
Copyright ? 2020 Elsevier Inc. All rights reserved.
KEYWORDS:
COVID-19; SARS-CoV; SARS-CoV-2; antiviral; coronavirus; vaccine
PMID:32289263DOI:10.1016/j.chom.2020.04.004
An Infectious cDNA Clone of SARS-CoV-2.
Xie X[SUP]1[/SUP], Muruato A[SUP]2[/SUP], Lokugamage KG[SUP]3[/SUP], Narayanan K[SUP]3[/SUP], Zhang X[SUP]4[/SUP], Zou J[SUP]4[/SUP], Liu J[SUP]3[/SUP], Schindewolf C[SUP]3[/SUP], Bopp NE[SUP]5[/SUP], Aguilar PV[SUP]6[/SUP], Plante KS[SUP]7[/SUP], Weaver SC[SUP]8[/SUP], Makino S[SUP]9[/SUP], LeDuc JW[SUP]10[/SUP], Menachery VD[SUP]11[/SUP], Shi PY[SUP]12[/SUP].
Author information
Abstract
The ongoing pandemic of COVID-19, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), underscores the urgency to develop experimental systems for studying this virus and identifying countermeasures. We report a reverse genetic system for SARS-CoV-2. Seven complimentary DNA (cDNA) fragments spanning the SARS-CoV-2 genome were assembled into a full-genome cDNA. RNA transcribed from the full-genome cDNA was highly infectious after electroporation into cells, producing 2.9 ? 10[SUP]6[/SUP] plaque-forming unit (PFU)/mL of virus. Compared with a clinical isolate, the infectious-clone-derived SARS-CoV-2 (icSARS-CoV-2) exhibited similar plaque morphology, viral RNA profile, and replication kinetics. Additionally, icSARS-CoV-2 retained engineered molecular markers and did not acquire other mutations. We generated a stable mNeonGreen SARS-CoV-2 (icSARS-CoV-2-mNG) by introducing this reporter gene into ORF7 of the viral genome. icSARS-CoV-2-mNG was successfully used to evaluate the antiviral activities of interferon (IFN). Collectively, the reverse genetic system and reporter virus provide key reagents to study SARS-CoV-2 and develop countermeasures.
Copyright ? 2020 Elsevier Inc. All rights reserved.
KEYWORDS:
COVID-19; SARS-CoV; SARS-CoV-2; antiviral; coronavirus; vaccine
PMID:32289263DOI:10.1016/j.chom.2020.04.004