A trans-complementation system for SARS-CoV-2 recapitulates authentic viral replication without virulence

Xianwen Zhang, Yang Liu, Jianying Liu, Adam L. Bailey, Kenneth S. Plante, Jessica A. Plante, Jing Zou, Hongjie Xia, Nathen E. Bopp, Patricia V. Aguilar, Ping Ren, Vineet D. Menachery, Michael S. Diamond, Scott C. Weaver, Xuping Xie, Pei Yong Shi

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish (US)
Pages (from-to)2229-2238.e13
Issue number8
StatePublished - Apr 15 2021


  • COVID-19
  • SARS-CoV-2
  • antiviral
  • coronavirus
  • diagnosis
  • vaccine

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology


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