@article{e9d2d18aef074472b4de8d38d04a559f,
title = "A trans-complementation system for SARS-CoV-2 recapitulates authentic viral replication without virulence",
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",
author = "Xianwen Zhang and Yang Liu and Jianying Liu and Bailey, {Adam L.} and Plante, {Kenneth S.} and Plante, {Jessica A.} and Jing Zou and Hongjie Xia and Bopp, {Nathen E.} and Aguilar, {Patricia V.} and Ping Ren and Menachery, {Vineet D.} and Diamond, {Michael S.} and Weaver, {Scott C.} and Xuping Xie and Shi, {Pei Yong}",
note = "Funding Information: We thank John Bilello from Gilead for providing Remdesivir and Zhiqiang An from the University of Texas Health Science at Houston for providing mAb14. We thank Q2 Solutions for the sponsored research agreement and scientific input toward this work. P.-Y.S. was supported by NIH grants AI134907 and UL1TR001439; awards from the Sealy & Smith Foundation, Kleberg Foundation, John S. Dunn Foundation, Amon G. Carter Foundation, Gilson Longenbaugh Foundation, and Summerfield Robert Foundation. M.S.D. was supported by R01 AI157155. V.D.M. was supported by NIH grants U19AI100625, R00AG049092, R24AI120942, and a STARs Award from the University of Texas System. S.C.W. was supported by NIH grant R24 AI120942. J.L. is supported by the postdoctoral fellowship from the McLaughlin Fellowship Endowment at UTMB. P.R. and X.X. were partially supported by the Sealy & Smith Foundation. X.Z. V.D.M. X.X. and P.-Y.S. conceived the study. X.Z. Y.L. J.L. A.L.B. K.S.P. J.A.P. J.Z. H.X. N.E.B. P.R. and X.X. performed the experiments. X.Z. Y.L. A.L.B. P.V.A. P.R. V.D.M. M.S.D. S.C.W. X.X. and P.-Y.S. analyzed the results. P.R. prepared the serum specimens. X.Z. Y.L. A.L.B. V.D.M. M.S.D. S.C.W. X.X. and P.-Y.S. wrote the manuscript. X.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. Funding Information: We thank John Bilello from Gilead for providing Remdesivir and Zhiqiang An from the University of Texas Health Science at Houston for providing mAb14. We thank Q2 Solutions for the sponsored research agreement and scientific input toward this work. P.-Y.S. was supported by NIH grants AI134907 and UL1TR001439 ; awards from the Sealy & Smith Foundation , Kleberg Foundation , John S. Dunn Foundation , Amon G. Carter Foundation , Gilson Longenbaugh Foundation , and Summerfield Robert Foundation . M.S.D. was supported by R01 AI157155 . V.D.M. was supported by NIH grants U19AI100625 , R00AG049092 , R24AI120942 , and a STARs Award from the University of Texas System . S.C.W. was supported by NIH grant R24 AI120942 . J.L. is supported by the postdoctoral fellowship from the McLaughlin Fellowship Endowment at UTMB . P.R. and X.X. were partially supported by the Sealy & Smith Foundation . Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",
year = "2021",
month = apr,
day = "15",
doi = "10.1016/j.cell.2021.02.044",
language = "English",
volume = "184",
pages = "2229--2238.e13",
journal = "Cell",
issn = "0092-8674",
number = "8",
}