Journal
SENSORS AND ACTUATORS B-CHEMICAL
Volume 312, Issue -, Pages -Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2020.127943
Keywords
Telomerase; Tetrahedral DNA nanoprobe; FRET-ON; Living cell imaging
Funding
- National Natural Science Foundation of China [81573390]
- National Key R&D Program of China [2018YFA0507204]
- Natural Science Foundation of Tianjin, China [18JCYBJC24800]
- Fundamental Research Funds for the Central Universities, Nankai University [63191519]
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Precise in situ monitoring of telomerase is highly informative in early cancer diagnosis. However, the complicated biological environment makes it difficult for conventional fluorescent probes to achieve effective cell penetration, stable delivery and accurate intracellular imaging. In this study, we design a functionalized tetrahedral DNA nanoprobe (TDNp) with fluorescence resonance energy transfer (FRET) feature to image telomerase in living cells. The proposed TDNp is composed of a DNA tetrahedral structure to improve stability and cellular permeability, a telomerase primer and a FRET-based molecular beacon (MB) to report fluorescent signal. In the presence of telomerase, the primer is extended and a free MB is recovered to trigger FRET response. The off-on FRET signal benefits accurate telomerase sensing with low false-positive feedback. By orienting the primer at the vertices of TDNp, we can sensitively detect telomerase with a detection limit of 35 HeLa cells. Furthermore, the TDNp could effectively distinguish cancer cells from normal cells, and is successfully applied to monitor the changes of telomerase activity in living cells. As a summary, the TDNp provides stable and reliable imaging of intracellular telomerase for cancer diagnosis.
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