Exploiting the potential of spherical PAM antenna for enhanced CRISPR-Cas12a: a paradigm shift toward a universal amplification-free nucleic acid test platform

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dc.contributor.authorDai, Jiahui
dc.contributor.authorWu, Beibei
dc.contributor.authorAi, Fengxiang
dc.contributor.authorYang, Zhugen
dc.contributor.authorLu, Yanyan
dc.contributor.authorZinian, Cai
dc.contributor.authorZeng, Kun
dc.contributor.authorZhang, Zhen
dc.date.accessioned2025-01-22T14:34:05Z
dc.date.available2025-01-22T14:34:05Z
dc.date.freetoread2025-01-22
dc.date.issued2025-01-09
dc.date.pubOnline2025-01-09
dc.description.abstractThe CRISPR-Cas12a system has shown tremendous potential for developing efficient biosensors. Albeit important, current CRISPR-Cas system-based diagnostic technologies (CRISPR-DX) highly rely on an additional preamplification procedure to obtain high sensitivity, inevitably leading to issues such as complicated assay workflow, cross-contamination, etc. Herein, a spherical protospacer-adjacent motif (PAM)-antenna-enhanced CRISPR-Cas12a system is fabricated for universal amplification-free nucleic acid detection with a detection limit of subfemtomolar. Meanwhile, the clinical detection capability of this sensor was further verified using gold-standard real-time quantitative polymerase chain reaction through <i>Mycobacterium tuberculosis</i> measurement, which demonstrated its good reliability for practical applications. Importantly, its excellent sensitivity is mainly ascribed to high efficiency of target search induced by a localized PAM-enriched microenvironment and improved catalytic activity of Cas12a (up to 4 folds). Our strategy provides some new insights for rapid and sensitive detection of nucleic acids in an amplification-free fashion.
dc.description.journalNameAnalytical Chemistry
dc.description.sponsorshipThis work was supported by the National Natural Science Foundation of China (grant nos. 22176075 and 22476072) and the Jiangsu Collaborative Innovation Center of Technology and Material of Water Treatment.
dc.description.sponsorshipLeverhulme Trust, National Natural Science Foundation of China.
dc.description.sponsorshipUKRI NERC Fellowship grant (NE/R013349/2).
dc.format.extentpp. 1236-1245
dc.format.mediumPrint-Electronic
dc.identifier.citationDai J, Wu B, Ai F, et al., (2025) Exploiting the potential of spherical PAM antenna for enhanced CRISPR-Cas12a: a paradigm shift toward a universal amplification-free nucleic acid test platform. Analytical Chemistry, Volume 97, Issue 2, January 2025, pp. 1236-1245
dc.identifier.eissn1520-6882
dc.identifier.elementsID562148
dc.identifier.issn0003-2700
dc.identifier.issueNo2
dc.identifier.urihttps://doi.org/10.1021/acs.analchem.4c04871
dc.identifier.urihttps://dspace.lib.cranfield.ac.uk/handle/1826/23417
dc.identifier.volumeNo97
dc.languageEnglish
dc.language.isoen
dc.publisherAmerican Chemical Society
dc.publisher.urihttps://pubs.acs.org/doi/10.1021/acs.analchem.4c04871
dc.rightsAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subject3401 Analytical Chemistry
dc.subject40 Engineering
dc.subject34 Chemical Sciences
dc.subjectEmerging Infectious Diseases
dc.subjectInfectious Diseases
dc.subjectBiotechnology
dc.subject3 Good Health and Well Being
dc.subjectAnalytical Chemistry
dc.subject3205 Medical biochemistry and metabolomics
dc.subject3401 Analytical chemistry
dc.subject4004 Chemical engineering
dc.titleExploiting the potential of spherical PAM antenna for enhanced CRISPR-Cas12a: a paradigm shift toward a universal amplification-free nucleic acid test platform
dc.typeArticle
dc.type.subtypeJournal Article
dcterms.dateAccepted2024-12-30

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