A structural reorganization-based catalytic hairpin assembly enabling small-molecule monitoring in living cells
Rui-Wen Wang1, Wei-Guo Yang1, Ming-Li Su1
1Key Laboratory of Luminescence Analysis and Molecular Sensing (Southwest University), Ministry of Education, Institute of Developmental Biology and Regenerative Medicine, College of Chemistry and Chemical Engineering, Southwest University Chongqing 400715 P. R. China wenbinliangasu@gmail.com.
Abstract:
Small-molecule drugs, constituting over 60% of FDA-approved therapeutics (2017-2022), present unresolved challenges relating to elucidating their intracellular mechanisms. We present a dual-strategy platform integrating "in silico aptamer affinity maturation" (ISAAM) and "structural reorganization-catalytic hairpin assembly" (SR-CHA). ISAAM computationally designs high-affinity aptamers, while SR-CHA eliminates undesired signals via energy-minimized conformational control, achieving a signal-to-background improvement over conventional CHA. This system enables ultrasensitive small-molecule monitoring in live cells, resolving traditional challenges of false positives and inefficiency. Demonstrated through intracellular imaging and kinetic studies, SR-CHA offers a robust tool for probing small-molecule interactions in biological systems, advancing drug discovery and diagnostic applications.
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