{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T04:18:40Z","timestamp":1791951520154,"version":"4.3.4"},"reference-count":25,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2021,12,29]],"date-time":"2021-12-29T00:00:00Z","timestamp":1640736000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R03AI163907"],"award-info":[{"award-number":["R03AI163907"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000054","name":"National Cancer Institute","doi-asserted-by":"publisher","award":["R01CA206592"],"award-info":[{"award-number":["R01CA206592"]}],"id":[{"id":"10.13039\/100000054","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R35GM130207"],"award-info":[{"award-number":["R35GM130207"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2022,2,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Enzymatic beacons, or E\u2010beacons, are 1\u2009:\u20091 bioconjugates of the nanoluciferase enzyme linked covalently at its C\u2010terminus to hairpin forming ssDNA equipped with a dark quencher. We prepared E\u2010beacons biocatalytically using HhC, the promiscuous Hedgehog C\u2010terminal protein\u2010cholesterol ligase. HhC attached nanoluciferase site\u2010specifically to mono\u2010sterylated hairpin oligonucleotides, called steramers. Three E\u2010beacon dark quenchers were evaluated: Iowa Black, Onyx\u2010A, and dabcyl. Each quencher enabled sensitive, sequence\u2010specific nucleic acid detection through enhanced E\u2010beacon bioluminescence upon target hybridization. We assembled prototype dabcyl\u2010quenched E\u2010beacons specific for SARS\u2010CoV\u20102. Targeting the E484 codon of the virus Spike protein, E\u2010beacons (80\u00d710\n                    <jats:sup>\u221212<\/jats:sup>\n                    \u2005M) reported wild\u2010type SARS\u2010CoV\u20102 nucleic acid at \u22651\u00d710\n                    <jats:sup>\u22129<\/jats:sup>\n                    \u2005M by increased bioluminescence of 8\u2010fold. E\u2010beacon prepared for the SARS\u2010CoV\u20102 E484K variant functioned with similar sensitivity. Both E\u2010beacons could discriminate their target from the E484Q mutation of the SARS\u2010CoV\u20102 Kappa variant. Along with mismatch specificity, E\u2010beacons are two to three orders of magnitude more sensitive than synthetic molecular beacons.\n                  <\/jats:p>","DOI":"10.1002\/cbic.202100594","type":"journal-article","created":{"date-parts":[[2021,12,10]],"date-time":"2021-12-10T11:37:20Z","timestamp":1639136240000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Enzymatic Beacons for Specific Sensing of Dilute Nucleic Acid**"],"prefix":"10.1002","volume":"23","author":[{"given":"Xiaoyu","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Chemistry, Binghamton University The State University of New York  4400 Vestal Parkway East Binghamton New York 13902 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Venubabu","family":"Kotikam","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Binghamton University The State University of New York  4400 Vestal Parkway East Binghamton New York 13902 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7649-0040","authenticated-orcid":false,"given":"Eriks","family":"Rozners","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Binghamton University The State University of New York  4400 Vestal Parkway East Binghamton New York 13902 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4826-0162","authenticated-orcid":false,"given":"Brian P.","family":"Callahan","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Binghamton University The State University of New York  4400 Vestal Parkway East Binghamton New York 13902 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,12,29]]},"reference":[{"key":"e_1_2_6_1_1","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.ES.2020.25.3.2000045"},{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2008457"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1021\/acssensors.0c01377"},{"key":"e_1_2_6_4_1","first-page":"2021","author":"Larremore D. B.","journal-title":"Sci. Adv."},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.2144\/btn-2020-0078"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abc6246"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1050-3862(98)00018-7"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0396-303"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/smll.201902248"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1021\/cb3002478"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1039\/C8AN00047F"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1039\/c1cc13495g"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST20200440"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-021-00878-8"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.bioconjchem.9b00550"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1039\/C8CC09729A"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.11.6171"},{"key":"e_1_2_6_18_1","unstructured":"Oligonucleotide sterylation for the prototype E-beacons Eb.1-3 was achieved by EDC coupling. After butanol extraction and RP-HPLC purification the final isolated yield of the sterylated oligo varied between 30\u201340\u2009%. The efficiency of oligonucleotide sterylation was improved significantly by switching to solid phase methods with a DNA synthesizer vide infra."},{"key":"e_1_2_6_19_1","unstructured":"We compared hybridization driven unquenching of with nuclease driven unquenching. Digestion of the hairpin component with DNASE-1 produced similar bioluminescence enhancement compared hybridization driven unquenching (Figure\u2005S4)."},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1177\/108705719900400206"},{"key":"e_1_2_6_21_1","unstructured":"https:\/\/covdb.stanford.edu\/page\/mutation-viewer."},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1039\/C6CC10032E"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciaa1382"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-021-02387-y"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.2144\/btn-2020-0157"}],"container-title":["ChemBioChem"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cbic.202100594","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cbic.202100594","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cbic.202100594","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T10:20:03Z","timestamp":1760264403000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/cbic.202100594"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,29]]},"references-count":25,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2022,2,16]]}},"alternative-id":["10.1002\/cbic.202100594"],"URL":"https:\/\/doi.org\/10.1002\/cbic.202100594","archive":["Portico"],"relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2021.08.30.458287","asserted-by":"object"}]},"ISSN":["1439-4227","1439-7633"],"issn-type":[{"value":"1439-4227","type":"print"},{"value":"1439-7633","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,29]]},"assertion":[{"value":"2021-10-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-12-29","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e202100594"}}