Abstract
Sensitive and specific sensing of intracellular telomerase is of significance for early cancer diagnosis and treatment. And, efficient signal amplification technology is a powerful tool to achieve low level biomarker detection. However, the delivery and release of amplification reagents in live cells is currently a major challenge to achieve sensitive intracellular detection of biomarkers. Here, we describe the development of a powerful in-cell amplification approach for label-free and highly sensitive detection of telomerase in live cells via zeolitic imidazolate framework-8 (ZIF-8)-mediated DNAzyme/light-up RNA aptamer transcription amplification cascades. The amplification reagents can be readily transported into and released in the target cells by ZIF-8 under intracellular acidic environment. Telomerase then activates DNAzyme to trigger the DNAzyme-aided cleavage reaction and T7 RNA polymerase-initiated light-up RNA aptamer transcription cycles for the yield of many light-up RNA aptamers, which associate with the non-fluorescent dye to recover significant fluorescence for achieving intracellular telomerase imaging with high sensitivity in a label-free approach. Moreover, the proposed strategy can discriminate telomerase level variations in different cells and shows promising potentials for anticancer drug screening, which makes such method a robust bioassay for early tumor-related diagnosis and therapeutic monitoring.
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