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Issue 29, 2015
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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

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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.

Graphical abstract: A highly sensitive fluorescent light-up probe for real-time detection of the endogenous protein target and its antagonism in live cells

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Publication details

The article was received on 04 May 2015, accepted on 18 Jun 2015 and first published on 19 Jun 2015


Article type: Communication
DOI: 10.1039/C5TB00819K
J. Mater. Chem. B, 2015,3, 5933-5937
  • Open access: Creative Commons BY-NC license
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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

    J. Geng, W. L. Goh, C. Zhang, D. P. Lane, B. Liu, F. Ghadessy and Y. N. Tan, J. Mater. Chem. B, 2015, 3, 5933
    DOI: 10.1039/C5TB00819K

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