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

doi:10.1016/j.cell.2021.02.044

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

Cell. 2021 Apr 15;184(8):2229-2238.e13. doi: 10.1016/j.cell.2021.02.044. Epub 2021 Feb 23.

Authors

Xianwen Zhang¹, Yang Liu¹, Jianying Liu², Adam L Bailey³, Kenneth S Plante⁴, Jessica A Plante⁴, Jing Zou¹, Hongjie Xia¹, Nathen E Bopp⁵, Patricia V Aguilar⁵, Ping Ren⁶, Vineet D Menachery², Michael S Diamond⁷, Scott C Weaver⁸, Xuping Xie⁹, Pei-Yong Shi¹⁰

Affiliations

¹ Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA.
² Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX, USA.
³ Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO, USA.
⁴ Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA.
⁵ Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA.
⁶ Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX, USA; Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA.
⁷ Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO, USA; Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA; Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO, USA.
⁸ Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX, USA; World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX, USA; Sealy Institute for Vaccine Sciences, University of Texas Medical Branch, Galveston, TX, USA.
⁹ Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX, USA. Electronic address: xuxie@UTMB.edu.
¹⁰ Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX, USA; Sealy Institute for Vaccine Sciences, University of Texas Medical Branch, Galveston, TX, USA; Sealy Center for Structural Biology & Molecular Biophysics, University of Texas Medical Branch, Galveston, TX, USA. Electronic address: peshi@UTMB.edu.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

A549 Cells
Animals
COVID-19 / virology*
Chlorocebus aethiops
Containment of Biohazards / methods*
Cricetinae
Genetic Complementation Test / methods
Genome, Viral
HEK293 Cells
Humans
Male
Mice
Mice, Transgenic
RNA, Viral
SARS-CoV-2* / genetics
SARS-CoV-2* / pathogenicity
SARS-CoV-2* / physiology
Vero Cells
Virulence
Virus Replication

Substances

RNA, Viral

Abstract

Publication types

MeSH terms

Substances

Grants and funding