Journal
NANOMEDICINE-NANOTECHNOLOGY BIOLOGY AND MEDICINE
Volume 32, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.nano.2020.102339
Keywords
Duplex-specific nuclease; G-quadruplex nanostring; Tandem rolling circle amplification; MicroRNA
Funding
- National Natural Science Foundation of China [21675072]
- Natural Science Foundation of Shandong Province [ZR2019MB068]
- Project of Shandong Province Higher Educational Science and Technology Program [KJ2018BZC043]
- Youth Innovation Team Project of Shandong Provincial Education Department
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A label-free and selective fluorescence platform for miRNA detection was developed using G-quadruplex nanostring and DSN-mediated RCA. The method showed excellent selectivity for perfectly matched DNA/RNA, with a detection limit of 1.019 fM for miR-21. This tool has potential applications in evaluating miR-21 levels in cell extracts for biomedical research and clinical diagnosis.
MicroRNA (miRNA) has emerged as a promising genetic marker for cancer diagnosis and therapy because its expression level is closely related to the progression of malignant diseases. Herein, a label-free and selective fluorescence platform was proposed for miRNA based on light-up G-quadruplex nanostring via duplex-specific nuclease (DSN) mediated tandem rolling circle amplification (RCA). First, a long DNA generated from upstream RCA was designed with the antisense sequences for miR-21 and downstream RCA primer. Upon recognizing miR-21, the resulting DNA RNA permitted DSN digestion and triggered downstream two-way RCA. and generation of abundant G-quadruplex nanostring binding with ZnPPIX for label-free fluorescent responses. In our strategy, the strong preference of DSN for perfectly matched DNA/RNA ensures its excellent selectivity. The developed method generated wide linear response with LOD of 1.019 fM. Additionally, the miR-21 levels in cell extracts have been evaluated, revealing the utility of this tool for biomedical research and clinical diagnosis. (C) 2020 Elsevier Inc. All rights reserved.
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