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Development of Fluorescence-Based Assays for Key Viral Proteins in the SARS-CoV-2 Infection Process and Lifecycle

Overview
Journal Int J Mol Sci
Publisher MDPI
Date 2024 Mar 13
PMID 38474097
Authors
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Abstract

Since the appearance of SARS-CoV-2 in 2019, the ensuing COVID-19 (Corona Virus Disease 2019) pandemic has posed a significant threat to the global public health system, human health, life, and economic well-being. Researchers worldwide have devoted considerable efforts to curb its spread and development. The latest studies have identified five viral proteins, spike protein (Spike), viral main protease (3CLpro), papain-like protease (PLpro), RNA-dependent RNA polymerase (RdRp), and viral helicase (Helicase), which play crucial roles in the invasion of SARS-CoV-2 into the human body and its lifecycle. The development of novel anti-SARS-CoV-2 drugs targeting these five viral proteins holds immense promise. Therefore, the development of efficient, high-throughput screening methodologies specifically designed for these viral proteins is of utmost importance. Currently, a plethora of screening techniques exists, with fluorescence-based assays emerging as predominant contenders. In this review, we elucidate the foundational principles and methodologies underpinning fluorescence-based screening approaches directed at these pivotal viral targets, hoping to guide researchers in the judicious selection and refinement of screening strategies, thereby facilitating the discovery and development of lead compounds for anti-SARS-CoV-2 pharmaceuticals.

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References
1.
Suh J, Kim H, Kim T . Development of a SARS-CoV-2-derived receptor-binding domain-based ACE2 biosensor. Sens Actuators B Chem. 2021; 334:129663. PMC: 7885701. DOI: 10.1016/j.snb.2021.129663. View

2.
Haga K, Takai-Todaka R, Sawada A, Katayama K . Luciferase-based quantification of membrane fusion induced by SARS-CoV-2 S protein. Genes Cells. 2022; 27(8):537-543. PMC: 9347681. DOI: 10.1111/gtc.12945. View

3.
Tian L, Qiang T, Liang C, Ren X, Jia M, Zhang J . RNA-dependent RNA polymerase (RdRp) inhibitors: The current landscape and repurposing for the COVID-19 pandemic. Eur J Med Chem. 2021; 213:113201. PMC: 7826122. DOI: 10.1016/j.ejmech.2021.113201. View

4.
Marx S, Mickolajczyk K, Craig J, Thomas C, Pfeffer A, Abell S . Observing inhibition of the SARS-CoV-2 helicase at single-nucleotide resolution. Nucleic Acids Res. 2023; 51(17):9266-9278. PMC: 10516658. DOI: 10.1093/nar/gkad660. View

5.
Abian O, Ortega-Alarcon D, Jimenez-Alesanco A, Ceballos-Laita L, Vega S, Reyburn H . Structural stability of SARS-CoV-2 3CLpro and identification of quercetin as an inhibitor by experimental screening. Int J Biol Macromol. 2020; 164:1693-1703. PMC: 7395220. DOI: 10.1016/j.ijbiomac.2020.07.235. View