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Updated: Aug 9, 2026
MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
Published on: October 7, 2025
Triply activated localized DNA circuit based on dual enzyme-guaranteed and analyte-stimulated strategy for robust
Yuanyuan Peng1, Xiaoyu Wang1, Wanling Cui2
1Shandong Key Laboratory of Applied Technology for Protein and Peptide Drugs, Institute of Biopharmaceutical Research, Liaocheng University, Liaocheng, 252000, China.
Background:
Localized catalytic hairpin assembly (LCHA) circuit has emerged as a potential tool for intracellular bioimaging, yet its application is limited by uncontrolled signal leakage, insufficient cell specificity, and unreliable readouts.
Results:
Herein, a triply activated LCHA (TA-LCHA) circuit was engineered by integrating an endogenous dual enzyme-guaranteed and analyte-stimulated strategy to enable robust tumor-cell-specific microRNA (miRNA) imaging. Initially, the TA-LCHA hairpin reactants were pre-blocked by incorporating the dual-enzyme recognition/cleavage site, inactivating their sensing function. Then, the miRNA sensing function of the blocked TA-LCHA reactants was specifically deblocked in tumor cells by two overexpressed enzymes, namely uracil DNA glycosylase (UDG) and apurinic/apyrimidinic endonuclease 1 (APE1). By virtue of the synergistic activation of endogenous UDG, APE1, and miRNA, this TA-LCHA circuit effectively reduced undesired signal leakage, achieving robust miRNA imaging with tumor cell specificity.
Significance:
The proposed TA-LCHA circuit represented a promising platform for precise biomolecules imaging in clinical diagnosis and biomedical applications.
