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A Highly Sensitive Fluorescent Light-up Probe for Real-time Detection of the Endogenous Protein Target and Its Antagonism in Live Cells

Lookup NU author(s): Dr Yen Nee Tan

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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0).


Abstract

Real-time detection and monitoring of cancer-related biomolecular interactions in live cells are of paramount importance for disease diagnostics and drug screening. Herein, we developed a target-specific fluorescent light-up probe for cellular detection of Mdm2, the key negative regulator of the p53 tumour suppressor protein. Conjugation of a uniquely designed fluorogen (TPECM) with aggregation induced-emission properties, to a specific p53-derived peptide (12.1Pep) targeting Mdm2, yielded a cell-permeable probe (TPECM–12.1Pep) with turn-on fluorescence properties for real-time live cell imaging of Mdm2. This specific light-up probe is almost non-fluorescent in its isolated state but is highly emissive upon binding to Mdm2, enabling quantitative detection of both Mdm2 and its antagonism. Using a model compound (Nutlin-3a), we demonstrate that the as-developed probes can be used to screen p53–Mdm2 inhibiting drug candidates, both in vitro and in cells. Furthermore, the probe activity can be accurately monitored in cells using a fluorescently activated cell sorting machine. These features will expedite research in the areas of drug discovery, clinical diagnostics and fundamental cell biology.


Publication metadata

Author(s): Geng J, Goh WLP, Zhang C, Lane DP, Liu B, Ghadessy FJ, Tan YN

Publication type: Article

Publication status: Published

Journal: Journal of Materials Chemistry B

Year: 2015

Volume: 3

Issue: 29

Pages: 5933-5937

Print publication date: 07/08/2015

Online publication date: 19/06/2015

Acceptance date: 18/06/2015

Date deposited: 07/04/2020

ISSN (print): 2050-750X

ISSN (electronic): 2050-7518

Publisher: Royal Society of Chemistry

URL: https://doi.org/10.1039/c5tb00819k

DOI: 10.1039/c5tb00819k


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Funding

Funder referenceFunder name
13302FG063
R-279-444-381

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