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Abstract
The strong correlation between cancer and telomerase activity has inspired the development of new strategies to evaluate telomerase activity. Here, a personal glucose meter (PGM) system that uses DNA-based machine amplification to detect telomerase in cancer cells is reported. In this assay, telomerase elongation products are amplified in the form of another type of product by a DNA-based machine. This process can only be activated by the hybridization of the extended telomerase substrate (TS) probe and the complementary primer in the presence of telomerase. The obtained products are then transformed to glucose-related signals via a three-component assay, which enables the simple use of a PGM to indirectly quantify the telomerase activity. The proposed method realizes sensitive telomerase activity detection down to 20 HeLa cells with a significantly enhanced dynamic range. Additionally, short telomerase elongation products, such as telomerase substrate probes with two repetitive sequences, that cannot be detected using the most widely used telomeric repeat amplification protocol assay were detected.
Highlights
Telomerase plays a vital role in cancer and ageing[1,2,3]
Encouraged by this, two assays using personal glucose meter (PGM) to evaluate telomerase activity have been reported recently, employing screen-printed gold electrodes (SPGE)[42], or mesoporous silica nanoparticles (MSN)[43]. They expand the application of PGM in telomerase activity detection undoubtedly, some shortcomings still exist for clinical use
We employed the synthetic telomerase elongation products telomerase substrate (TS) + 2R, TS + 3R, and TS + 4R, which correspond to the TS probe extended by two, three and four telomeric repetitive sequences of (TTA GGG) to evaluate the feasibility of the proposed method
Summary
Telomerase plays a vital role in cancer and ageing[1,2,3]. Telomerase is a reverse transcriptase composed of a protein and a template RNA component. Among the strategies used to detect telomerase activity, polymerase chain reaction (PCR)-based telomeric repeat amplification protocol (TRAP) is the most frequently used[3,14,15,16] This method is extraordinary sensitive for the detection of telomerase activity at the single-cell level, quantification is difficult, and precise temperature cycling control is required. We exploited a widely commercialized PGM to develop a novel method for the detection of telomerase activity in cell extracts based on a DNA-based machine that further improves the dynamic range and sensitivity. In this strategy, a tailored TS probe is the “track” of the machine and is first extended by telomerase. This method possesses the potential to be developed into a point of care (POC) diagnostic device for telomerase detection
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