PT  - JOURNAL ARTICLE
AU  - Xianwen Zhang
AU  - Yang Liu
AU  - Jianying Liu
AU  - Adam L. Bailey
AU  - Kenneth S. Plante
AU  - Jessica A. Plante
AU  - Jing Zou
AU  - Hongjie Xia
AU  - Nathen Bopp
AU  - Patricia Aguilar
AU  - Ping Ren
AU  - Vineet D. Menachery
AU  - Michael S. Diamond
AU  - Scott C. Weaver
AU  - Xuping Xie
AU  - Pei-Yong Shi
TI  - A <em>trans</em>-complementation system for SARS-CoV-2
AID  - 10.1101/2021.01.16.426970
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.01.16.426970
4099  - http://biorxiv.org/content/early/2021/01/19/2021.01.16.426970.short
4100  - http://biorxiv.org/content/early/2021/01/19/2021.01.16.426970.full
AB  - The biosafety level-3 (BSL-3) requirement to culture severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a bottleneck for research and countermeasure development. 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 a deletion of ORF3 and envelope gene, 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. The results suggest that the trans-complementation platform can be safely used at BSL-2 laboratories for research and countermeasure development.Competing Interest StatementX.Z., X.X., and P.-Y.S. have filed a patent on the trans-complementation system of SARS-CoV-2. M.S.D. is a consultant for Inbios, Vir Biotechnology, NGM Biopharmaceuticals, and Carnival Corporation, and on the Scientific Advisory Boards of Moderna and Immunome. The Diamond laboratory has received unrelated funding support in sponsored research agreements from Moderna, Vir Biotechnology, and Emergent BioSolutions.