{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T06:01:50Z","timestamp":1790661710301,"version":"4.1.0"},"reference-count":45,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T00:00:00Z","timestamp":1790553600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T00:00:00Z","timestamp":1790553600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Small Methods"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>The programmability of\u00a0CRISPR\u2010Cas12a\u00a0has enabled transformative advances in nucleic acid detection; however, extending its activation to non\u2010nucleic\u2010acid targets remains highly desirable. Here, we report a universal split\u2010activator strategy that enables programmable Cas12a activation in response to a small molecule and a protein. In this approach, a canonical activator is divided into two fragments (p1 and p2), such that Cas12a activation occurs only upon their simultaneous assembly. We integrate this mechanism with aptamer\u2010mediated conformation switching, wherein target binding destabilizes an aptamer\u2010p2 duplex, releasing p2 to complete the activator complex and trigger Cas12a nuclease activity. Using this design, we demonstrate selective recognition of the small\u2010molecule metabolite adenosine and the protein biomarker prostate\u2010specific antigen (PSA), with high specificity against structurally related controls. Signal generation is achieved using fluorescent DNA nanoclusters (DFN\u20101), which undergo fluorescence decrease upon Cas12a\u2010mediated degradation. The system exhibits concentration\u2010dependent responses for both targets. Together, this work establishes a modular framework for coupling aptamer\u2010based molecular recognition to programmable Cas12a activation, expanding the functional scope of CRISPR systems beyond nucleic acids and enabling new opportunities in CRISPR\u2010Cas12a\u2010mediated detection.<\/jats:p>","DOI":"10.1002\/smtd.71075","type":"journal-article","created":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T05:24:16Z","timestamp":1790659456000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Split Activator Assembly via Aptamer Binding Enables CRISPR\u2010Cas12a Activation With Non\u2010Nucleic Acid Targets"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-2402-1346","authenticated-orcid":false,"given":"Ram J.","family":"Tharu","sequence":"first","affiliation":[{"name":"Department of Chemistry University At Albany State University of New York  Albany New York 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