tetano
Editor, Senior Moderator
Cell
. 2021 Feb 23;S0092-8674(21)00233-6.
doi: 10.1016/j.cell.2021.02.044. Online ahead of print.
A trans-complementation system for SARS-CoV-2 recapitulates authentic viral replication without virulence
Xianwen Zhang[SUP] 1 [/SUP], Yang Liu[SUP] 1 [/SUP], Jianying Liu[SUP] 2 [/SUP], Adam L Bailey[SUP] 3 [/SUP], Kenneth S Plante[SUP] 4 [/SUP], Jessica A Plante[SUP] 4 [/SUP], Jing Zou[SUP] 1 [/SUP], Hongjie Xia[SUP] 1 [/SUP], Nathen E Bopp[SUP] 5 [/SUP], Patricia V Aguilar[SUP] 5 [/SUP], Ping Ren[SUP] 6 [/SUP], Vineet D Menachery[SUP] 2 [/SUP], Michael S Diamond[SUP] 7 [/SUP], Scott C Weaver[SUP] 8 [/SUP], Xuping Xie[SUP] 9 [/SUP], Pei-Yong Shi[SUP] 10 [/SUP]
Affiliations
Abstract
The biosafety level 3 (BSL-3) requirement to culture severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a bottleneck for research. Here, we report a trans-complementation system that produces single-round infectious SARS-CoV-2 that recapitulates authentic viral replication. We demonstrate that the single-round infectious SARS-CoV-2 can be used at BSL-2 laboratories for high-throughput neutralization and antiviral testing. The trans-complementation system consists of two components: a genomic viral RNA containing ORF3 and envelope gene deletions, as well as mutated transcriptional regulator sequences, and a producer cell line expressing the two deleted genes. Trans-complementation of the two components generates virions that can infect naive cells for only one round but does not produce wild-type SARS-CoV-2. Hamsters and K18-hACE2 transgenic mice inoculated with the complementation-derived virions exhibited no detectable disease, even after intracranial inoculation with the highest possible dose. Thus, the trans-complementation platform can be safely used at BSL-2 laboratories for research and countermeasure development.
Keywords: COVID-19; SARS-CoV-2; antiviral; coronavirus; diagnosis; vaccine.
. 2021 Feb 23;S0092-8674(21)00233-6.
doi: 10.1016/j.cell.2021.02.044. Online ahead of print.
A trans-complementation system for SARS-CoV-2 recapitulates authentic viral replication without virulence
Xianwen Zhang[SUP] 1 [/SUP], Yang Liu[SUP] 1 [/SUP], Jianying Liu[SUP] 2 [/SUP], Adam L Bailey[SUP] 3 [/SUP], Kenneth S Plante[SUP] 4 [/SUP], Jessica A Plante[SUP] 4 [/SUP], Jing Zou[SUP] 1 [/SUP], Hongjie Xia[SUP] 1 [/SUP], Nathen E Bopp[SUP] 5 [/SUP], Patricia V Aguilar[SUP] 5 [/SUP], Ping Ren[SUP] 6 [/SUP], Vineet D Menachery[SUP] 2 [/SUP], Michael S Diamond[SUP] 7 [/SUP], Scott C Weaver[SUP] 8 [/SUP], Xuping Xie[SUP] 9 [/SUP], Pei-Yong Shi[SUP] 10 [/SUP]
Affiliations
- PMID: 33691138
- DOI: 10.1016/j.cell.2021.02.044
Abstract
The biosafety level 3 (BSL-3) requirement to culture severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a bottleneck for research. Here, we report a trans-complementation system that produces single-round infectious SARS-CoV-2 that recapitulates authentic viral replication. We demonstrate that the single-round infectious SARS-CoV-2 can be used at BSL-2 laboratories for high-throughput neutralization and antiviral testing. The trans-complementation system consists of two components: a genomic viral RNA containing ORF3 and envelope gene deletions, as well as mutated transcriptional regulator sequences, and a producer cell line expressing the two deleted genes. Trans-complementation of the two components generates virions that can infect naive cells for only one round but does not produce wild-type SARS-CoV-2. Hamsters and K18-hACE2 transgenic mice inoculated with the complementation-derived virions exhibited no detectable disease, even after intracranial inoculation with the highest possible dose. Thus, the trans-complementation platform can be safely used at BSL-2 laboratories for research and countermeasure development.
Keywords: COVID-19; SARS-CoV-2; antiviral; coronavirus; diagnosis; vaccine.