One-pot detection of telomerase activity with high sensitivity and specificity <i>via</i> RNA FRET probes and RNase H-assisted signal cycling amplification

Wang, Honghong; Wang, Hui; Jia, Yuting; Li, Zhengping

doi:10.1039/c9ra01816f

Cited by 1 publication

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References 31 publications

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“…It had been reported that the G-quadruplex was more stable in the presence of K + than that formed in the presence of Na + . 25 Therefore, we studied the ThT fluorescence change after the incubation of ThT with the TS primer and telomerase extract in telomerase extension reaction buffer in the presence of different concentrations of K + . As shown in Supplemental Figures S2A and S2B, the ThT fluorescence intensity increased on increasing the concentration of K + due to the fact that formation of the G-quadruplex structure was induced and stabilized by potassium, while ThT showed a relatively weak fluorescence emission in the absence of K + .…”

Section: Resultsmentioning

confidence: 99%

A label- and enzyme-free fluorescence assay based on thioflavin T–induced G-quadruplexes for the detection of telomerase activity

Chen

2023

Journal of Chemical Research

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A label- and enzyme-free fluorescence assay based on thioflavin T–induced G-quadruplexes is developed to sensitively and specifically detect telomerase activity. Thioflavin T has a dual role as an efficient inducer and fluorescent probe, and the incorporation of thioflavin T into the thioflavin T–induced G-quadruplexes results in an intense fluorescence enhancement. In the presence of thioflavin T and K+, G-quadruplexes are formed by elongation of the telomerase substrate primer that is catalyzed by telomerase extracted from cancer cells. Thus, the telomerase activity in cancer cell extracts can be evaluated by measuring the thioflavin T fluorescence. More importantly, thioflavin T can specifically recognize and bind to G-quadruplexes, whereas it cannot recognize single- and double-stranded DNAs, which leads to the thioflavin T–based fluorescence assay exhibiting a reduced background and improved signal-to-noise ratio. As a result, the proposed assay has the linear range from 5 to 200 HeLa cells and the detection limit is 34 HeLa cells, which holds great potential for use in the detection of telomerase activity and the diagnosis of cancer.

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Section: Resultsmentioning

confidence: 99%

A label- and enzyme-free fluorescence assay based on thioflavin T–induced G-quadruplexes for the detection of telomerase activity

Chen

2023

Journal of Chemical Research

View full text Add to dashboard Cite

show abstract

One-pot detection of telomerase activity with high sensitivity and specificity via RNA FRET probes and RNase H-assisted signal cycling amplification

Abstract: A one-pot method is developed for the detection of telomerase activity via RNA FRET probes and RNase H-assisted signal cycling amplification.

Cited by 1 publication

References 31 publications

A label- and enzyme-free fluorescence assay based on thioflavin T–induced G-quadruplexes for the detection of telomerase activity

A label- and enzyme-free fluorescence assay based on thioflavin T–induced G-quadruplexes for the detection of telomerase activity

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