ArticleNature communications2026
Moderate-temperature DNA cleavage activity of TtAgo activated by dCTP/TthSSB for one-step isothermal microRNAs detection.
Article in Nature communications, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Thermus thermophilus Argonaute (TtAgo) is a DNA-guided endonuclease promising for gene editing and molecular diagnostics. However, its high-temperature dependence (65-85 °C) restricts its utility in moderate-temperature scenarios. Here we show that combining deoxycytidine triphosphate (dCTP) with the single-stranded DNA-binding protein from T. thermophilus (TthSSB) enables robust TtAgo activity within 37-60 °C. Mechanistic studies reveal that dCTP increases TtAgo's conformational flexibility, while TthSSB promotes substrate recruitment, synergistically driving efficient DNA cleavage. Leveraging this mechanism, we develop ERCB-TtAgo (exponential amplification reaction by dCTP/TthSSB-activated TtAgo), a one-step isothermal diagnostic platform for microRNA detection. It achieves femtomolar sensitivity within 40 min and accurately distinguishes hepatocellular carcinoma patients from controls in a 151-sample clinical cohort, matching RT‑qPCR performance. Our work not only surmounts a key limitation of TtAgo but also offers an approach for modulating nuclease activity, paving the way for intelligent biosensing platforms with broad applicability.
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