4.7 Article

Target-triggered assembly of functional G-quadruplex DNAzyme nanowires for sensitive detection of miRNA in lung tissues

Journal

SENSORS AND ACTUATORS B-CHEMICAL
Volume 373, Issue -, Pages -

Publisher

ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2022.132689

Keywords

Nanowires; MicroRNA; Thioflavin T; Cancer cells; Nonsmall-cell lung carcinoma (NSCLC) patients

Funding

  1. National Natural Science Foundation of China
  2. Project of Shan- dong Province Higher Educational Science and Technology Program
  3. Award for Team Leader Program of Taishan Scholars of Shandong Province, China
  4. [21735003]
  5. [21605098]
  6. [J18KA105]

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In this study, a novel method was developed for sensitive detection of miRNA in lung tissues by target-triggered assembly of G-quadruplex DNAzyme nanowires. This method exhibited high sensitivity, a large dynamic range, and the ability to distinguish single-base mutations, holding promising applications in biomedical analysis and cancer prognosis.
MicroRNAs (miRNAs) are important biomarkers for multiple diseases including cancers, neurodegenerative diseases, and cardiovascular diseases. Accurate detection of miRNAs is imperative for early diagnosis and biomedical research. Herein, we demonstrate target-triggered assembly of G-quadruplex DNAzyme nanowires for sensitive detection of miRNAs in lung tissues by integrating primer generation-mediated rolling circle amplifi-cation (PG-RCA) with primer exchange reaction (PER). Target miRNA hybridizes with the circular template to initiate PG-RCA, producing numerous primers. The obtained primers then react with the catalytic hairpin DNA template to trigger PER, producing the polymer G-quadruplex DNAzyme nanowires. Eventually, thioflavin T (ThT) as a fluorescence indicator selectively interacts with the G-quadruplex DNAzyme to generate an amplified signal. This assay is capable of detecting miRNA over a large dynamic range (up to 6 orders of magnitude) and it exhibits high sensitivity with a limit of detection (LOD) of 61.34 aM. Moreover, this assay displays excellent specificity and it can distinguish even a single-base mutation in miR-486-5p. Furthermore, this method can quantify the miR-486-5p concentration in different types of cells, and it enables the identification of differen-tially expressed miR-486-5p in lung tissues of normal persons and nonsmall-cell lung carcinoma (NSCLC) pa-tients, holding promising applications in biomedical analysis and cancer prognosis.

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