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

A highly sensitive fluorescent light-up probe for real-time detection of the endogenous protein target and its antagonism in live cells
Title:
A highly sensitive fluorescent light-up probe for real-time detection of the endogenous protein target and its antagonism in live cells
Other Titles:
Journal of Materials Chemistry B
Publication Date:
19 June 2015
Citation:
J. Mater. Chem. B, 2015,3, 5933-5937
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.
License type:
http://creativecommons.org/licenses/by-nc/4.0/
Funding Info:
The authors would like to acknowledge the Agency for Science, Technology and Research (A*STAR), Singapore, for the financial support under the JCO CDA grant 13302FG063 and Singapore National Research Foundation (R-279-444-381).
Description:
ISSN:
2050-750X
2050-7518
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