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Updated: May 6, 2026
Combining QD-FRET and Microfluidics to Monitor DNA Nanocomplex Self-Assembly in Real-Time
Published on: August 26, 2009
Nicking endonuclease-propelled DNA nanoassemblies enable cross-validated trimodal biosensing with ultrahigh
Xu Li1, Yashu Wei2, Peiyuan Li1
1Key Laboratory of Optic-electric Chemo/Biosensing and Molecular Recognition(Guangxi Minzu University), Education Department of Guangxi Zhuang Autonomous Region, School of Chemistry and Chemical Engineering, Guangxi Minzu University, Nanning, 530006, China.
None:
Current biosensing platforms face fundamental limitations in reliability and environmental adaptability due to single-signal output constraints, including susceptibility to false positives, poor anti-interference capability, and limited applicability in complex matrices. To overcome these critical bottlenecks, we develop a nicking endonuclease-propelled trimodal biosensing platform that integrates electrochemical (EC), colorimetric (CL), and photothermal (PTM) detection through an innovative rolling circle amplification (RCA) mechanism. The system's core innovation lies in the synergistic integration of nicking endonuclease-assisted cyclic amplification with DNA nanoassembly technology, enabling the construction of nanoflower-shaped nucleic acid structures that provide substantial signal amplification and cross-validation capability. Materially, we engineer a novel AuNPs/Zr-MOF/GDY nanocomposite with exceptional electron transfer capability, enlarged surface area, and abundant active sites, addressing the critical limitation of poor electrical conductivity in MOF-based sensing platforms. Mechanistically, the platform features nicking endonuclease-initiated RCA amplification creating dual-output DNA nanostructures, DNAzyme walker-driven cascade reactions for exponential signal enhancement, and triple-mode output providing built-in validation against false signals. It demonstrates exceptional CD17 detection performance across a 0.0001-100,000 pM linear range with ultra-low detection limits of 31.7 aM (EC), 11.2 fM (CL), and 10.3 fM (PTM) (S/N = 3). Successful human serum validation showing 97.7-106.3 % recovery confirms strong clinical utility for early thalassemia screening. This work provides a generalizable framework for next-generation biosensing design, offering fundamental solutions to reliability challenges through multi-mode cross-validation, enzyme-driven signal amplification, and robust nanoarchitectures adaptable to various biomarker detection scenarios.
