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However, current methods typically involve an amplification process, require substantial time, and are susceptible to aerosol contamination. Herein, we introduce a NanoLock-powered, amplification-free assay based on the type III-E clustered regularly interspaced short palindromic repeats (CRISPR)\u2013CRISPR-associated system for rapid, highly sensitive, and specific RNA diagnostics. This innovative platform, designated CRISPR-guided caspase (Craspase)\u2013NanoLock\u2013Csx30 (CNC), harmoniously integrates the precise protease activity of Craspase with the remarkable luminescent sensitivity of NanoLock, creating a novel and streamlined approach for RNA detection. The CNC platform exhibited exceptional sensitivity in detecting severe acute respiratory syndrome coronavirus-2 N\u00a0gene RNA through the integration of three guide RNAs, achieving a detection limit of 250 fM in just 10 min\u00a0without amplification. Preliminary studies further revealed the platform's extended diagnostic potential for detecting influenza A virus and human immunodeficiency virus. These findings collectively establish the CNC platform as an appealing tool for infectious disease detection and significantly broaden the scope of CRISPR-based diagnostic applications.<\/jats:p>","DOI":"10.1093\/nar\/gkaf907","type":"journal-article","created":{"date-parts":[[2025,9,18]],"date-time":"2025-09-18T19:52:22Z","timestamp":1758225142000},"source":"Crossref","is-referenced-by-count":3,"title":["A NanoLock-enabled, Craspase-based strategy for highly sensitive RNA detection"],"prefix":"10.1093","volume":"53","author":[{"given":"Yumeng","family":"Xiao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Microbial Technology, Shandong University , 72 Binhai Road , Qingdao 266237 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junyu","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Microbial Technology, Shandong University , 72 Binhai Road , Qingdao 266237 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