Academic Journal
Click-chemistry-mediated modulation of CRISPR-Cas12a activity through activator modification.
| Τίτλος: | Click-chemistry-mediated modulation of CRISPR-Cas12a activity through activator modification. |
|---|---|
| Συγγραφείς: | Fu R; Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, 310016, China; College of Biosystems Engineering and Food Science, Zhejiang Key Laboratory of Agro-Food Resources and High-Value Utilization, Zhejiang University, Hangzhou, 310058, China., Zhu C; Department of Laboratory Medicine, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian, 350207, China; Fujian Clinical Research Center for Laboratory Medicine of Immunology, The First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian, 350005, China., Hou J; Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, 310016, China; College of Biosystems Engineering and Food Science, Zhejiang Key Laboratory of Agro-Food Resources and High-Value Utilization, Zhejiang University, Hangzhou, 310058, China., Wang Z; Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, 310016, China; College of Biosystems Engineering and Food Science, Zhejiang Key Laboratory of Agro-Food Resources and High-Value Utilization, Zhejiang University, Hangzhou, 310058, China., Xianyu Y; Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, 310016, China; College of Biosystems Engineering and Food Science, Zhejiang Key Laboratory of Agro-Food Resources and High-Value Utilization, Zhejiang University, Hangzhou, 310058, China. Electronic address: xianyu19@zju.edu.cn. |
| Πηγή: | Biosensors & bioelectronics [Biosens Bioelectron] 2026 Dec 01; Vol. 313, pp. 119144. Date of Electronic Publication: 2026 Aug 22. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Elsevier Advanced Technology Country of Publication: England NLM ID: 9001289 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4235 (Electronic) Linking ISSN: 09565663 NLM ISO Abbreviation: Biosens Bioelectron Subsets: MEDLINE |
| Imprint Name(s): | Publication: Oxford : Elsevier Advanced Technology Original Publication: [Barking, Essex, England] : Elsevier Applied Science, 1989- |
| Ιατρικοί όροι (MeSH): | Click Chemistry*/methods , CRISPR-Cas Systems*/genetics , CRISPR-Associated Proteins*/chemistry , CRISPR-Associated Proteins*/genetics , CRISPR-Associated Proteins*/metabolism , Bacterial Proteins*/chemistry , Bacterial Proteins*/genetics , Endodeoxyribonucleases*/chemistry , Endodeoxyribonucleases*/genetics , Biosensing Techniques*, Azides/chemistry ; DNA/chemistry ; DNA/genetics ; Alkynes/chemistry ; Cyclooctanes/chemistry ; Cycloaddition Reaction ; Molecular Docking Simulation |
| Περίληψη: | Chemical modification strategies offer a promising route for spatiotemporal regulation of CRISPR-Cas12a activity in molecular diagnostics. However, existing methods involve CRISPR RNA with photolabile groups that suffer from complexity and RNA instability. To address these limitations, we report a simple and robust strategy using dibenzocyclooctyne (DBCO)-mediated click chemistry to modulate CRISPR-Cas12a activity. The copper-free strain-promoted azide-alkyne cycloaddition reaction enables CRISPR-Cas12a modulation with low toxicity, biocompatibility, and high selectivity. Utilizing azide-modified non-target DNA strand sequences at different locations to react with DBCO, we show that DBCO-modified activators can regulate CRISPR-Cas12a cleavage in three distinct states: maintain, enhance, and suppress. Mechanistic studies through cleavage kinetics and molecular docking reveal that the regulatory outcome depends on the modification position, protospacer adjacent motif composition, and DBCO concentration. We further employ asymmetric polymerase chain reaction to generate azide-modified DNA for click-chemistry-mediated modulation of CRISPR-Cas12a. This strategy could be a promising tool for regulating CRISPR-Cas12a activity in molecular diagnostics. (Copyright © 2026 Elsevier B.V. All rights reserved.) |
| Competing Interests: | Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. |
| Contributed Indexing: | Keywords: CRISPR-Cas12a; Cas12a activity modulation; Click chemistry; DNA activator; Dibenzocyclooctyne |
| Substance Nomenclature: | 0 (Azides) 9007-49-2 (DNA) 0 (CRISPR-Associated Proteins) 0 (Alkynes) 0 (Cyclooctanes) EC 3.1.- (Cas12a protein) 0 (Bacterial Proteins) EC 3.1.- (Endodeoxyribonucleases) |
| Entry Date(s): | Date Created: 20260825 Date Completed: 20260904 Latest Revision: 20260904 |
| Update Code: | 20260905 |
| DOI: | 10.1016/j.bios.2026.119144 |
| PMID: | 42641278 |
| Βάση Δεδομένων: | MEDLINE |
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