Rapid screening of SARS-CoV-2 inhibitors via ratiometric fluorescence of RBD–ACE2 complexes in living cells by competitive binding

Liu, Miao; Zhou, Wei; Yan, Chunyu; Zhang, Yuebin; Qiao, Qinglong; Zhou, Xuelian; Chen, Yingzhu; Wang, Guangying; Guo, Zhendong; Li, Jun; Piao, Hailong; Pan, Xin; Yan, Ming; Zhao, Weijie; Li, Guohui; Li, Yueqing; Xu, Zhaochao

doi:10.1016/j.apsb.2022.05.033

Cited by 8 publications

(5 citation statements)

References 6 publications

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“…The development of optical imaging techniques has allowed research into targeted drugs to be performed at a high resolution and to be monitored in vivo in real time 33 , 34 , 35 . In order to generate useful data for these applications, it is necessary to generate high-quality images at the subcellular, cellular, and animal scales and to employ advanced image quantification tools to process complex information.…”

Section: Advanced Fluorescence Imaging Technologies and Analytical Toolsmentioning

confidence: 99%

Enhanced optical imaging and fluorescent labeling for visualizing drug molecules within living organisms

Sun,

Zhao,

et al. 2024

Acta Pharmaceutica Sinica B

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Section: Advanced Fluorescence Imaging Technologies and Analytical Toolsmentioning

confidence: 99%

Enhanced optical imaging and fluorescent labeling for visualizing drug molecules within living organisms

Sun,

Zhao,

et al. 2024

Acta Pharmaceutica Sinica B

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“…Fluorescent imaging of ACE2 has also been used for evaluating novel agents for inhibiting RBD binding to ACE2. Again, using SNAP-and Halo-tag-labeled ACE2 and RBD, respectively, inhibitors were screened on living cells, as the fluorescent signal from Halo-RBD will not stain cells docking when ACE2 is blocked [22]. A second method for screening ACE2 inhibitors involves the fluorescence signal generated by protein-protein interactions [23].…”

Section: Fluorescent Imagingmentioning

confidence: 99%

Molecular Imaging of ACE2 Expression in Infectious Disease and Cancer

Li,

Hasson,

Daggumati

et al. 2023

Viruses

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Angiotensin-converting enzyme 2 (ACE2) is a cell-surface receptor that plays a critical role in the pathogenesis of SARS-CoV-2 infection. Through the use of ligands engineered for the receptor, ACE2 imaging has emerged as a valuable tool for preclinical and clinical research. These can be used to visualize the expression and distribution of ACE2 in tissues and cells. A variety of techniques including optical, magnetic resonance, and nuclear medicine contrast agents have been developed and employed in the preclinical setting. Positron-emitting radiotracers for highly sensitive and quantitative tomography have also been translated in the context of SARS-CoV-2-infected and control patients. Together this information can be used to better understand the mechanisms of SARS-CoV-2 infection, the potential roles of ACE2 in homeostasis and disease, and to identify potential therapeutic modulators in infectious disease and cancer. This review summarizes the tools and techniques to detect and delineate ACE2 in this rapidly expanding field.

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“…Fluorescence labeling and live-cell tracking techniques enable dynamic visualization of protein function with high spatial and temporal resolution. Protein self-labeling tags (SNAP-tag, Halo-tag as commonly used ones) can singly link small molecule dyes to proteins of interest, which were widely used for live-cell protein imaging to monitor protein–protein or protein–molecule interaction, including single-virus visual tracking. − The biggest advantage of it is that various small molecule probes (brightness, photostability, color-tunable et al) can be selected for different experiment requirements. − …”

Section: Introductionmentioning

confidence: 99%

Live-Cell Imaging to Resolve Salt-Induced Liquid–Liquid Phase Separation of FUS Protein by Dye Self-Labeling

Zhang,

Xu,

Yan

et al. 2023

Chemical & Biomedical Imaging

Self Cite

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The aggregation of fusion in sarcoma (FUS) in the cytoplasm and nucleus is a pathological feature of Amyotrophic lateral sclerosis (ALS) and Frontotemporal Dementia (FTD). Genetic mutations, abnormal protein synthesis, environmental stress, and aging have all been implicated as causative factors in this process. Salt ions are essential to many physiological processes in the body, and the imbalance of them is an important environmental stress factor in cells. However, their effect on liquid−liquid phase separation (LLPS) of FUS proteins in living cells is not well understood. Here, we map the various salt-induced LLPS of FUS in living cells by genetically coding and self-labeling FUS with organic dyes. The brightness and photostability of the dyes enable long-term imaging to track the mechanism of the assembly and disappearance of FUS phase separation. The FUS protein showed a better phase separation tendency under 0.3 M salt stimulation, and there was a large amount of FUS shuttling from the nucleus to the cytoplasm. At this concentration, various salt solutions displayed different effects on the phase separation of FUS protein, following the Hofmeister effects. We further observed that the assembly of FUS droplets underwent a process of rapid formation of small droplets, plateaus, and mutual fusion. Strikingly, The CsCl-stimulated FUS droplets were not completely reversible after washing, and some solid-like granules remained in the nucleus. Taken together, these results help broaden our understanding of the LLPS of FUS proteins in cellular stress responses.

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Rapid screening of SARS-CoV-2 inhibitors via ratiometric fluorescence of RBD–ACE2 complexes in living cells by competitive binding

Cited by 8 publications

References 6 publications

Enhanced optical imaging and fluorescent labeling for visualizing drug molecules within living organisms

Enhanced optical imaging and fluorescent labeling for visualizing drug molecules within living organisms

Molecular Imaging of ACE2 Expression in Infectious Disease and Cancer

Live-Cell Imaging to Resolve Salt-Induced Liquid–Liquid Phase Separation of FUS Protein by Dye Self-Labeling

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