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12 16, 2022;3 (4): 101802.

Protocol for characterizing the inhibition of SARS-CoV-2 infection by a protein of interest in cultured cells.

Xinyuan Lai, Hui Zhuang, Tong Li, Kuanhui Xiang
Author Information
  1. Xinyuan Lai: Department of Microbiology and Infectious Disease Center, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China; Peking University-YHLO Joint Laboratory for Molecular Diagnostics of Infectious Diseases, Peking University, Beijing 100191, China.
  2. Hui Zhuang: Department of Microbiology and Infectious Disease Center, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China; Peking University-YHLO Joint Laboratory for Molecular Diagnostics of Infectious Diseases, Peking University, Beijing 100191, China.
  3. Tong Li: Department of Microbiology and Infectious Disease Center, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China; Peking University-YHLO Joint Laboratory for Molecular Diagnostics of Infectious Diseases, Peking University, Beijing 100191, China.
  4. Kuanhui Xiang: Department of Microbiology and Infectious Disease Center, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China; Peking University-YHLO Joint Laboratory for Molecular Diagnostics of Infectious Diseases, Peking University, Beijing 100191, China. Electronic address: kxiang@bjmu.edu.cn.
PMID: 36345374 DOI: 10.1016/j.xpro.2022.101802

Abstract

Here, we present a protocol to characterize the antiviral ability of a protein of interest to SARS-CoV-2 infection in cultured cells, using MUC1 as an example. We use SARS-CoV-2 ΔN trVLP system, which utilizes transcription and replication-competent SARS-CoV-2 virus-like particles lacking nucleocapsid gene. We describe the optimized procedure to analyze protein interference of viral attachment and entry into cells, and qRT-PCR-based quantification of viral infection. The protocol can be applied to characterize more antiviral candidates and clarify their functioning stage. For complete details on the use and execution of this protocol, please refer to Lai et al. (2022).

Keywords

Cell Biology Cell-based Assays Microbiology Molecular Biology Protein Bbiochemistry

References

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MeSH Term

Humans
COVID-19
SARS-CoV-2
Nucleocapsid
Cell Line
Antiviral Agents

Chemicals

Antiviral Agents
Journal Article Research Support, Non-U.S. Gov't
Article has an altmetric score of 1

Word Cloud

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