Journal
ANALYTICAL CHEMISTRY
Volume 94, Issue 11, Pages 4874-4880Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c00583
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Funding
- National Key R&D Program of China [2020YFA0211200, 2021YFA1200403, 2018YFE0206900]
- National Natural Science Foundation of China [22090050, 21974128, 21874121, 52003257]
- Joint NSFC-ISF Research Grant Program [22161142020]
- Hubei Provincial Natural Science Foundation of China [2020CFA037]
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This study introduced a dual signal output detection strategy based on cell-cycle synchronization to precisely detect telomerase activities using a new AIEgen probe SSNB. Experimental results demonstrated the effectiveness of this strategy in indicating telomerase activity in tumor cells, with the highest activity observed in the S phase.
By maintaining the telomere lengths, telomerase can make the tumor cells avoid the apoptosis, thus, achieving the cell immortalization. In the past, a series of telomerase detection systems have been developed through utilizing the unique characteristic of telomerase extended primer. However, fluctuation of telomerase activity, along with the cell cycle progression, leads to ambiguous detection results. Here, we reported a dual signal output detection strategy based on cell-cycle synchronization for precisely detecting telomerase activities by using a new AIEgen probe SSNB. Experimental and simulating calculation results demonstrated that positively charged SSNB could interact with DNA through the electrostatic interaction and pi-pi interaction, as well as the hydrogen bonds. The aggregation of SSNB caused by the extended template strand primer (TP) could be observed in tumor cells, thus, indicating the telomerase activity in various cell lines. Furthermore, after cell cycle synchronization, it was found that the telomerase activity in the S phase was the highest, no matter from the fluorescence intensity or the ROS generation situation. Dual signal outputs of SSNB verified the significance and necessity of cell-cycle synchronization detection for telomerase activity. This strategy could open a new window for the biotargets of which activity is variational in time dimension.
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