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Herein, we report an enzyme-free and indicator-free electrochemical biosensor for microRNA-21 based on allosteric switching-triggered electrocatalyst recruitment and hydrogen evolution reaction (HER) readout. The sensing interface comprises a hairpin probe DNA that undergoes target-induced allosteric switching upon hybridization with microRNA-21, thereby exposing a streptavidin-binding domain for recruitment of the HER electrocatalyst FLN-Pt@SA. This target-triggered interfacial assembly converts molecular recognition into a catalyst-amplified HER response, using H2O as an abundant electrochemical substrate rather than conventional redox indicators or mediators. The resulting biosensor generates a stable milliampere-level signal and enables quantitative detection of microRNA-21 over a wide concentration range from 100 aM to 100 nM, with a detection limit of 34 aM. The biosensor also exhibits good selectivity against non-target and mismatched miRNA sequences, as well as satisfactory reproducibility, stability, and recovery in serum samples. This work establishes an enzyme-free, indicator-free HER-based signal-transduction strategy that couples nucleic acid recognition with electrocatalytic amplification for electrochemical biosensing.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c04584","type":"journal-article","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T15:01:17Z","timestamp":1789052477000},"page":"27612-27625","source":"Crossref","is-referenced-by-count":0,"title":["Allosteric\nSwitching-Triggered Electrocatalyst Recruitment\nEnables Indicator-Free, Milliampere-Level Hydrogen Evolution Readout\nfor MicroRNA-21 Detection"],"prefix":"10.1021","volume":"98","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-9923-9453","authenticated-orcid":true,"given":"Yifan","family":"Chen","sequence":"first","affiliation":[{"name":"Minnan Normal University , , ,","place":["Zhangzhou, P. R. China, 363000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zehao","family":"Wang","sequence":"additional","affiliation":[{"name":"Minnan Normal University , , ,","place":["Zhangzhou, P. R. China, 363000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5173-5918","authenticated-orcid":true,"given":"Xuan","family":"Zheng","sequence":"additional","affiliation":[{"name":"Minnan Normal University , , ,","place":["Zhangzhou, P. R. China, 363000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Gao","sequence":"additional","affiliation":[{"name":"Minnan Normal University , , ,","place":["Zhangzhou, P. R. China, 363000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiancong","family":"Ni","sequence":"additional","affiliation":[{"name":"Minnan Normal University , , ,","place":["Zhangzhou, P. R. China, 363000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8952-9925","authenticated-orcid":true,"given":"Qingxiang","family":"Wang","sequence":"additional","affiliation":[{"name":"Minnan Normal University , , ,","place":["Zhangzhou, P. R. China, 363000"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,10]]},"reference":[{"issue":"7","key":"2026092208080580000_cit1","doi-asserted-by":"publisher","first-page":"4049","DOI":"10.1021\/acs.analchem.4c05841","article-title":"N-Deficient B-Doped g-C3N4\/CdS Heterojunction-Based PEC-FL Biosensor Assisted\nby CRISPR-Cas12a System for Ultrasensitive Determination of microRNA","volume":"97","author":"Du","year":"2025","journal-title":"Anal. 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