Authors
Zhen Yu, Jin Wang, Yacong An, Jing Wu, Shuangshuang Du, Yujing Wu, Jiajian Wang, Jiandong Zhang
Published in
Analytical methods : advancing methods and applications. Nov 04, 2026. Epub Nov 04, 2026.
Abstract
MicroRNA-122 (miR-122) is a liver-enriched miRNA closely associated with hepatic disorders, making its sensitive detection crucial for liver disease-related biomarker analysis. Herein, we report a magnetic bead-assisted duplex-specific nuclease (DSN) cleavage strategy coupled with CRISPR/Cas12a fluorescence biosensing for sensitive miR-122 detection. A biotinylated DNA probe containing both a miR-122 recognition region and a Cas12a activator region was immobilized on streptavidin-modified magnetic beads. Upon target hybridization, DSN selectively cleaved the RNA/DNA duplex, releasing the DNA activator while enabling miR-122 recycling. After magnetic separation, the released activator triggered Cas12a-mediated trans-cleavage of a fluorophore-quencher-labeled ssDNA reporter, generating a fluorescence turn-on signal. Under optimized conditions, the biosensor showed a linear response from 100 fM to 10 nM, with a detection limit of 85 fM. The assay exhibited high specificity against mismatched sequences and other miRNAs, and achieved satisfactory recoveries (93.0-95.3%, RSD < 8%) in 10% human serum. This magnetic bead-assisted DSN/CRISPR-Cas12a fluorescence biosensor provides a simple, sensitive, and specific platform for miR-122 detection, holding promise for liver disease-related diagnostic applications.
PMID:
42980643
Bibliographic data and abstract were imported from PubMed on 04 Nov 2026.
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