Facile, Fast-Responsive, and Photostable Imaging of Telomerase Activity in Living Cells with a Fluorescence Turn-On Manner

Abstract

In situ detecting and monitoring intracellular telomerase activity is significant for cancer diagnosis. In this work, we report a facile and fast-responsive bioprobe for in situ detection and imaging of intracellular telomerase activity with superior photostability. After transfected into living cells, quencher group labeled TS primer (QP) can be extended in the presence of intracellular telomerase. Positive charged TPE-Py molecules (AIE dye) will bind to the primer as well as extension repeated units, producing a telomerase activity-related turn-on fluorescence signal. By incorporating positive charged AIE dye and substrate oligonucleotides, in situ light-up imaging and detection of intracellular telomerase activity were achieved. This strategy exhibits good performance for sensitive in situ tracking of telomerase activity in living cells. The practicality of this facile and fast-responsive telomerase detection method was demonstrated by using it to distinguish tumor cells from normal cells and to monitor the change of telomerase activity during treatment with antitumor drugs, which shows its potential in clinical diagnostic and therapeutic monitoring.

Advances in the detection of telomerase activity using isothermal amplification

“…Besides the above AIEgens-based in vitro telomerase activity assays, imaging of telomerase activity with a turn-on manner in living cells has been also reported by Lou et al 82. They made use of a yellow-emissive AIE dye (TPE-Py) for in situ monitoring intracellular telomerase activity ( Figure 10 ).…”

Sensing telomerase: From in vitro detection to in vivo imaging

“…With the introduction of novel nanomaterials, the in vivo imaging of telomerase in living cells may be achieved. 49,52,53,56 Both in vitro and in vivo telomerase assays make a great contribution to biomedical research and clinical diagnostics.…”

“…8). 49 The TPE-Py is a kind of aggregation-induced emission (AIE) dye with weak fluorescence in the separation state but intense fluorescence in the aggregation state. 50 The telomerase substrate is labelled with a quencher.…”

“…In addition to the fluorescent imaging methods, 49,52,53 Kuang demonstrated the use of in situ Raman scattering spectroscopy for the intracellular telomerase assay (Fig. 11).…”

Ultrasmall Organic Nanoparticles with Aggregation-Induced Emission and Enhanced Quantum Yield for Fluorescence Cell Imaging

“…[13][14][15][16][17][18] Among the various optical imaging agents, particle-based imaging agents [5][6][7] are of great interest since organic molecular dyes often suffers from short retention time and are subject to photo-bleaching. 19, 20 However, most particle-based fluorophores are composed of inorganic species, even some toxic heavy metal cations, of which the potential cytotoxicity effects have greatly impaired their application.…”