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Updated: Jul 17, 2026
MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
Published on: October 7, 2025
Programmable DNA nanowires mediated dispersion-to-localization catalytic hairpin assembly for high-contrast miRNA
Ziheng Tang1, Kefan Zheng1, Bochen Liu2
1Zhejiang Provincial Key Laboratory of Medical Genetics, Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Abstract:
Accurate profiling of microRNAs (miRNAs) is often limited by the severe background leakage of conventional enzyme-free DNA amplification. Herein, we develop a 1D DNA nanowire-mediated dispersion-to-localization catalytic hairpin assembly (DL-CHA) platform for the highly sensitive detection of miRNAs. Methodologically, responsive hairpins are physically isolated on spatially dispersed nanowire scaffolds to rigorously suppress target-independent interactions. Upon specific recognition of miR-21, a pseudo-intramolecular cascade is triggered, dynamically cross-linking the scaffolds into a highly fluorescent DNA ladder. As a result, this DL-CHA platform yields an exceptional signal-to-background ratio of 12.4, a broad linear range of 0.05 to 250 nM, and an ultra-sensitive limit of detection of 50 pM without the need for exogenous enzymes. Furthermore, the rigid nanowire architecture ensures robust nuclease resistance and enables highly efficient, transfection-free internalization for the in situ imaging of endogenous miRNAs in living cells. Clinically, the platform accurately discriminated colorectal cancer patients from healthy controls using non-invasive liquid biopsies of human serum. This paradigm offers a robust, low-background molecular tool for precision cancer diagnostics.
