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Circular DNA aptamers, with their dramatically enhanced biological and structural stability, hold great potential as new\u2010generation DNA\u2010based anticoagulants. However, the functional selection of circular aptamers and large\u2010scale synthesis of them remains a substantial challenge. In this study, we explored a binding\u2010site directed selection strategy to enrich circular DNA aptamers that can target the catalytic domain of FXIa and inhibit its function. An asymmetrical dumbbell structured DNA library with self\u2010shielding primer\u2010binding site was utilized for selection. This approach has resulted in the identification of a circular DNA aptamer, named FICAPT1, which demonstrates a high binding affinity (\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>d<\/jats:sub>\n                    =20\u2005nM), notable stability (with a half\u2010life of 16\u2005h in human plasma) and outstanding anticoagulation activity (with significantly prolonged aPTT), showing great promise as a novel anticoagulant. Moreover, a refined version of FICAPT1, named as CC\u2010FICAPT1, was quantitatively synthesized (~2\u2005mg\/mL) by taking advantage of the pre\u2010designed structure and high cyclization efficiency of click chemistry. The approach presented in this study enables the efficient generation of functional circular DNA aptamers and allows for their synthesis in a satisfactory scale, making it adaptable for the production of various circular aptamers for therapeutic applications.\n                  <\/jats:p>","DOI":"10.1002\/chem.202404372","type":"journal-article","created":{"date-parts":[[2025,1,17]],"date-time":"2025-01-17T02:23:07Z","timestamp":1737080587000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Binding\u2010Site Directed Selection and Large\u2010Scale Click\u2010Synthesis of a Coagulation Factor XIa\u2010Inhibiting Circular DNA Aptamer"],"prefix":"10.1002","volume":"31","author":[{"given":"Jie","family":"Wang","sequence":"first","affiliation":[{"name":"School of Food and Biological Engineering Hefei University of Technology  Hefei 230009 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuefei","family":"Zhou","sequence":"additional","affiliation":[{"name":"School of Food and Biological Engineering Hefei University of Technology  Hefei 230009 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Long","family":"Sun","sequence":"additional","affiliation":[{"name":"School of Food and Biological Engineering Hefei University of Technology  Hefei 230009 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Food and Biological Engineering Hefei University of Technology  Hefei 230009 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Qu","sequence":"additional","affiliation":[{"name":"School of Food and Biological Engineering Hefei University of Technology  Hefei 230009 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4656-2864","authenticated-orcid":false,"given":"Yu","family":"Mao","sequence":"additional","affiliation":[{"name":"School of Food and Biological Engineering Hefei University of Technology  Hefei 230009 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Zheng","sequence":"additional","affiliation":[{"name":"School of Food and Biological Engineering Hefei University of Technology  Hefei 230009 China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,1,23]]},"reference":[{"key":"e_1_2_8_1_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature06797"},{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0049-3848(16)30363-2"},{"key":"e_1_2_8_3_1","unstructured":"\u00a0"},{"key":"e_1_2_8_4_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2009-05-219915"},{"key":"e_1_2_8_5_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM200003093421004"},{"key":"e_1_2_8_6_1","unstructured":"\u00a0"},{"key":"e_1_2_8_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2022.121021"},{"key":"e_1_2_8_8_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jmedchem.1c00613"},{"key":"e_1_2_8_9_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jmedchem.1c00056"},{"key":"e_1_2_8_10_1","unstructured":"\u00a0"},{"key":"e_1_2_8_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2018.04.007"},{"key":"e_1_2_8_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.2200121"},{"key":"e_1_2_8_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/344467a0"},{"key":"e_1_2_8_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrd3141"},{"key":"e_1_2_8_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/346818a0"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-02055-x"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.thromres.2017.06.015"},{"key":"e_1_2_8_18_1","unstructured":"\u00a0"},{"key":"e_1_2_8_19_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.9b09884"},{"key":"e_1_2_8_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.2c09093"},{"key":"e_1_2_8_21_2","doi-asserted-by":"publisher","DOI":"10.1039\/D0SC02279A"},{"key":"e_1_2_8_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-019-0090-6"},{"key":"e_1_2_8_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.9b11490"},{"key":"e_1_2_8_24_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.7b04547"},{"key":"e_1_2_8_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.8b03442"},{"key":"e_1_2_8_26_2","author":"Guo S. 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