{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T19:41:05Z","timestamp":1791661265151,"version":"4.3.4"},"reference-count":43,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T00:00:00Z","timestamp":1774483200000},"content-version":"vor","delay-in-days":7,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2022YFC2304304"],"award-info":[{"award-number":["2022YFC2304304"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Science and Technology Innovation Talent Plan of Hubei Province","award":["2023DJC136"],"award-info":[{"award-number":["2023DJC136"]}]},{"DOI":"10.13039\/501100003819","name":"Natural Science Foundation of Hubei Province","doi-asserted-by":"publisher","award":["2025AFB825"],"award-info":[{"award-number":["2025AFB825"]}],"id":[{"id":"10.13039\/501100003819","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,3,19]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Current one-pot clustered regularly interspaced short palindromic repeats diagnostics are limited by the cis-cleavage activity of Cas nucleases, which leads to amplicon degradation during amplification. Here, we report a streamlined strategy that overcomes this limitation. By integrating a bipartite split-crRNA into Cas12a (SCas12a), we separate target recognition from PAM dependency and completely eliminate cis-cleavage while preserving robust trans-cleavage. This strategy is broadly applicable for one-pot testing, compatible with recombinase polymerase amplification, RT\u2013RPA, and loop-mediated isothermal amplification, as well as multiple Cas12a orthologs, including As, Lb, and Ct Cas12a. Moreover, the SCas12a accelerates one-pot testing with 100\u20131000-fold improved sensitivity and achieves &amp;gt;10-fold reduction in time-to-signal, enabling detection of targets at attomolar levels within 30 min. Additionally, it provides single-base resolution with up to 91-fold selectivity. The system has been successfully applied to detect HPV16, SARS-CoV-2, and TP53 SNPs in clinical samples. Together, we have developed a PAM-independent and cis-cleavage-free one-pot Cas12a assay, which holds strong potential for point-of-care diagnostics.<\/jats:p>","DOI":"10.1093\/nar\/gkag267","type":"journal-article","created":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T11:12:11Z","timestamp":1774523531000},"source":"Crossref","is-referenced-by-count":8,"title":["Elimination of\n                    <i>cis<\/i>\n                    -cleavage in CRISPR diagnostics for one-pot rapid nucleic acid detection"],"prefix":"10.1093","volume":"54","author":[{"given":"Wenhao","family":"Yin","sequence":"first","affiliation":[{"name":"State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University , Hubei 430042 ,","place":["China"]},{"name":"School of Life Science and Technology, Wuhan Polytechnic University , Hubei 430023 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhili","family":"Jin","sequence":"additional","affiliation":[{"name":"Department of Cardiology, Zhongnan Hospital of Wuhan University , Hubei 430071 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingyuan","family":"Jiang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University , Hubei 430042 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuqi","family":"Jin","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University , Hubei 430042 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinping","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocatalysis and Enzyme 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,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xianhua","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University , Hubei 430042 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2148-3596","authenticated-orcid":false,"given":"Yi","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University , Hubei 430042 ,","place":["China"]},{"name":"BravoVax Co., Ltd., Wuhan , Hubei 430075 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,3,26]]},"reference":[{"key":"2026032607120508100_B1","doi-asserted-by":"publisher","first-page":"eabd5393","DOI":"10.1126\/sciadv.abd5393","article-title":"Test 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