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Here, we report a DNA nanoruler\u2010assisted linearization strategy that mechanically regulates LCNA conformation by employing a rigid DNA tetrahedral framework (DTF) dimer as a nanoruler. This approach anchors the target termini at a predefined nanoscale spacing (\u223c10\u201315\u00a0nm), imposing defined spatial constraints that stretch and pre\u2010open secondary structures, leading to linearization and enhanced site accessibility. In homogeneous assays, this strategy delivers robust signal enhancement across LCNAs with diverse structures. Integrated into an electrochemical platform, it achieves LODs of 1 copy\/\u00b5L for the E and N genes and 10 copies\/\u00b5L for ORF1ab, with linear ranges of 10\u201310\n                    <jats:sup>5<\/jats:sup>\n                    copies\/\u00b5L for E gene and 10\n                    <jats:sup>2<\/jats:sup>\n                    \u201310\n                    <jats:sup>5<\/jats:sup>\n                    copies\/\u00b5L for both N gene and ORF1ab from SARS\u2010CoV\u20102. Furthermore, when incorporated into a fluorescence chip, this method enables high\u2010throughput analysis of circulating cfDNA (ALU115) for prostate cancer discrimination with an AUC of 90.35%. Collectively, we establish a DNA nanoruler\u2010based conformational regulation strategy as a structure\u2010tolerant approach for high\u2010performance LCNA detection in both electrochemical and fluorescent modes.\n                  <\/jats:p>","DOI":"10.1002\/anie.1686086","type":"journal-article","created":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T12:12:55Z","timestamp":1787227975000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["DNA Nanoruler\u2010Assisted Linearization for Long\u2010Chain Nucleic Acids Analysis"],"prefix":"10.1002","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7709-0433","authenticated-orcid":false,"given":"Xi","family":"Yu","sequence":"first","affiliation":[{"name":"Renji Branch of National Center for Translational Medicine Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine Institute of Molecular Medicine Shanghai Key Laboratory for Nucleic Acid 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Regulation Shanghai Institute of Measurement and Testing Technology Co., Ltd  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0021-5135","authenticated-orcid":false,"given":"Xiurong","family":"Yang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Synergistic Chem\u2010Bio Synthesis School of Chemistry and Chemical Engineering New Cornerstone Science Laboratory Frontiers Science Center for Transformative Molecules Zhangjiang Institute for Advanced Study The Institute of Artificial Intelligence and National Center for Translational Medicine Shanghai Jiao Tong University  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7171-7338","authenticated-orcid":false,"given":"Chunhai","family":"Fan","sequence":"additional","affiliation":[{"name":"Renji Branch of National Center for Translational Medicine Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine Institute of Molecular Medicine Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine Renji Hospital Shanghai Jiao Tong University School of Medicine  Shanghai China"},{"name":"State Key Laboratory of Synergistic Chem\u2010Bio Synthesis School of Chemistry and Chemical Engineering New Cornerstone Science Laboratory Frontiers Science Center for Transformative Molecules Zhangjiang Institute for Advanced Study The Institute of Artificial Intelligence and National Center for Translational Medicine Shanghai Jiao Tong University  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7505-2727","authenticated-orcid":false,"given":"Xiaolei","family":"Zuo","sequence":"additional","affiliation":[{"name":"Renji Branch of National Center for Translational Medicine Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine Institute of Molecular Medicine Shanghai Key Laboratory for Nucleic 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