دورية أكاديمية
Broad-spectrum enzymatic inhibition of CRISPR-Cas12a.
| العنوان: | Broad-spectrum enzymatic inhibition of CRISPR-Cas12a. |
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| المؤلفون: | Knott GJ; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA., Thornton BW; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA., Lobba MJ; Department of Chemistry, University of California, Berkeley, CA, USA., Liu JJ; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA., Al-Shayeb B; Department of Plant and Microbial Biology, University of California, Berkeley, CA, USA., Watters KE; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA., Doudna JA; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA. doudna@berkeley.edu.; Department of Chemistry, University of California, Berkeley, CA, USA. doudna@berkeley.edu.; Molecular Biophysics & Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA. doudna@berkeley.edu.; Gladstone Institutes, San Francisco, CA, USA. doudna@berkeley.edu.; Howard Hughes Medical Institute, University of California, Berkeley, CA, USA. doudna@berkeley.edu.; Innovative Genomics Institute, University of California, Berkeley, CA, USA. doudna@berkeley.edu. |
| المصدر: | Nature structural & molecular biology [Nat Struct Mol Biol] 2019 Apr; Vol. 26 (4), pp. 315-321. Date of Electronic Publication: 2019 Apr 01. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
| اللغة: | English |
| بيانات الدورية: | Publisher: Nature Pub. Group Country of Publication: United States NLM ID: 101186374 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1545-9985 (Electronic) Linking ISSN: 15459985 NLM ISO Abbreviation: Nat Struct Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: New York : Nature Pub. Group, c2004- |
| مواضيع طبية MeSH: | CRISPR-Cas Systems/*physiology, Adaptive Immunity/genetics ; Adaptive Immunity/physiology ; CRISPR-Cas Systems/genetics ; DNA Cleavage ; Endoribonucleases/genetics ; Endoribonucleases/metabolism ; Gene Editing/methods ; Protein Multimerization/genetics ; Protein Multimerization/physiology |
| مستخلص: | Cas12a is a bacterial RNA-guided nuclease used widely for genome editing and, more recently, as a molecular diagnostic. In bacteria, Cas12a enzymes can be inhibited by bacteriophage-derived proteins, anti-CRISPRs (Acrs), to thwart clustered regularly interspaced short palindromic repeat (CRISPR) adaptive immune systems. How these inhibitors disable Cas12a by preventing programmed DNA cleavage is unknown. We show that three such inhibitors (AcrVA1, AcrVA4 and AcrVA5) block Cas12a activity via functionally distinct mechanisms, including a previously unobserved enzymatic strategy. AcrVA4 and AcrVA5 inhibit recognition of double-stranded DNA (dsDNA), with AcrVA4 driving dimerization of Cas12a. In contrast, AcrVA1 is a multiple-turnover inhibitor that triggers cleavage of the target-recognition sequence of the Cas12a-bound guide RNA to irreversibly inactivate the Cas12a complex. These distinct mechanisms equip bacteriophages with tools to evade CRISPR-Cas12a and support biotechnological applications for which multiple-turnover enzymatic inhibition of Cas12a is desirable. |
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| معلومات مُعتمدة: | United States HHMI Howard Hughes Medical Institute |
| المشرفين على المادة: | EC 3.1.- (Endoribonucleases) |
| تواريخ الأحداث: | Date Created: 20190403 Date Completed: 20191125 Latest Revision: 20230425 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC6449189 |
| DOI: | 10.1038/s41594-019-0208-z |
| PMID: | 30936531 |
| قاعدة البيانات: | MEDLINE |
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Full Text Finder | تدمد: | 1545-9985 |
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| DOI: | 10.1038/s41594-019-0208-z |