{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T09:35:25Z","timestamp":1786440925235,"version":"build-2736575974"},"reference-count":37,"publisher":"Oxford University Press (OUP)","issue":"15","license":[{"start":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:00:00Z","timestamp":1786406400000},"content-version":"vor","delay-in-days":1,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["U23A20265"],"award-info":[{"award-number":["U23A20265"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32530084"],"award-info":[{"award-number":["32530084"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003995","name":"Anhui Provincial Natural Science Foundation","doi-asserted-by":"publisher","award":["2408085MC072"],"award-info":[{"award-number":["2408085MC072"]}],"id":[{"id":"10.13039\/501100003995","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012226","name":"Fundamental Research Funds for the Central Universities","doi-asserted-by":"publisher","award":["PA2025GDGP0026"],"award-info":[{"award-number":["PA2025GDGP0026"]}],"id":[{"id":"10.13039\/501100012226","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system has become a powerful genome-editing tool that uses RNA\u2013DNA pairing to cleave target DNA with protospacer adjacent motif (PAM) sequences. While its primary function is well-studied, secondary activities remain poorly understood, causing unintended off-target effects. This study reports for the first time that Cas9 specifically cleaves the 5\u2032 overhang of the non-target strand (NTS) in target double-stranded DNA. This specific cleavage requires an additional PAM element at the NTS 5\u2032 region, is mediated by Cas9\u2019s RuvC domain, and is regulated by the HNH domain. It depends on the exact positioning of the NTS 5\u2032 end, but not on the overhang homopolymer sequence or overhang length. Adequate single-guide RNA\u2013DNA complementarity is also essential. This discovery potentially advances our understanding of Cas9\u2019s enzymatic versatility to enhance genome-editing precision and efficacy and offers new nucleic acid detection strategies. Based on this cleavage, we developed a sensitive assay for Severe Acute Respiratory Syndrome Coronavirus 2 pseudovirus down to 2.4 copies \u03bcL\u22121, demonstrated extremely high sensitivity in diagnostic applications.<\/jats:p>","DOI":"10.1093\/nar\/gkag795","type":"journal-article","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T11:58:41Z","timestamp":1785412721000},"source":"Crossref","is-referenced-by-count":0,"title":["Specific cleavage of 5\u2032 overhangs of non-target strand by Cas9 activated by target DNA binding"],"prefix":"10.1093","volume":"54","author":[{"given":"Shaokang","family":"Lv","sequence":"first","affiliation":[{"name":"School of Food and Biological Engineering, Hefei University of Technology , Hefei 230009 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Food and Biological Engineering, Hefei University of Technology , Hefei 230009 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Provincial International Science and Technology Cooperation Base for Major Metabolic Diseases and Nutritional Interventions , Hefei 230009 ,","place":["China"]},{"name":"Intelligent Interconnected Systems Laboratory of Anhui Province, Hefei University of Technology , Hefei 230009 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,8,11]]},"reference":[{"key":"2026081105000498000_B1","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1093\/genetics\/28.6.491","article-title":"Mutations of bacteria from virus sensitivity to virus resistance","volume":"28","author":"Luria","year":"1943","journal-title":"Genetics"},{"key":"2026081105000498000_B2","doi-asserted-by":"publisher","first-page":"1390","DOI":"10.1128\/JB.01412-07","article-title":"Phage response to CRISPR-encoded resistance in Streptococcus thermophilus","volume":"190","author":"Deveau","year":"2008","journal-title":"J 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