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SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation
Characteristic properties of type III CRISPR-Cas systems include recognition of target RNA and the subsequent induction of a multifaceted immune response. This involves sequence-specific cleavage of the target RNA and production of cyclic oligoadenylate (cOA) molecules. Here we report that an expose...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8376896/ https://www.ncbi.nlm.nih.gov/pubmed/34413302 http://dx.doi.org/10.1038/s41467-021-25337-5 |
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author | Steens, Jurre A. Zhu, Yifan Taylor, David W. Bravo, Jack P. K. Prinsen, Stijn H. P. Schoen, Cor D. Keijser, Bart J. F. Ossendrijver, Michel Hofstra, L. Marije Brouns, Stan J. J. Shinkai, Akeo van der Oost, John Staals, Raymond H. J. |
author_facet | Steens, Jurre A. Zhu, Yifan Taylor, David W. Bravo, Jack P. K. Prinsen, Stijn H. P. Schoen, Cor D. Keijser, Bart J. F. Ossendrijver, Michel Hofstra, L. Marije Brouns, Stan J. J. Shinkai, Akeo van der Oost, John Staals, Raymond H. J. |
author_sort | Steens, Jurre A. |
collection | PubMed |
description | Characteristic properties of type III CRISPR-Cas systems include recognition of target RNA and the subsequent induction of a multifaceted immune response. This involves sequence-specific cleavage of the target RNA and production of cyclic oligoadenylate (cOA) molecules. Here we report that an exposed seed region at the 3′ end of the crRNA is essential for target RNA binding and cleavage, whereas cOA production requires base pairing at the 5′ end of the crRNA. Moreover, we uncover that the variation in the size and composition of type III complexes within a single host results in variable seed regions. This may prevent escape by invading genetic elements, while controlling cOA production tightly to prevent unnecessary damage to the host. Lastly, we use these findings to develop a new diagnostic tool, SCOPE, for the specific detection of SARS-CoV-2 from human nasal swab samples, revealing sensitivities in the atto-molar range. |
format | Online Article Text |
id | pubmed-8376896 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-83768962021-09-02 SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation Steens, Jurre A. Zhu, Yifan Taylor, David W. Bravo, Jack P. K. Prinsen, Stijn H. P. Schoen, Cor D. Keijser, Bart J. F. Ossendrijver, Michel Hofstra, L. Marije Brouns, Stan J. J. Shinkai, Akeo van der Oost, John Staals, Raymond H. J. Nat Commun Article Characteristic properties of type III CRISPR-Cas systems include recognition of target RNA and the subsequent induction of a multifaceted immune response. This involves sequence-specific cleavage of the target RNA and production of cyclic oligoadenylate (cOA) molecules. Here we report that an exposed seed region at the 3′ end of the crRNA is essential for target RNA binding and cleavage, whereas cOA production requires base pairing at the 5′ end of the crRNA. Moreover, we uncover that the variation in the size and composition of type III complexes within a single host results in variable seed regions. This may prevent escape by invading genetic elements, while controlling cOA production tightly to prevent unnecessary damage to the host. Lastly, we use these findings to develop a new diagnostic tool, SCOPE, for the specific detection of SARS-CoV-2 from human nasal swab samples, revealing sensitivities in the atto-molar range. Nature Publishing Group UK 2021-08-19 /pmc/articles/PMC8376896/ /pubmed/34413302 http://dx.doi.org/10.1038/s41467-021-25337-5 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Steens, Jurre A. Zhu, Yifan Taylor, David W. Bravo, Jack P. K. Prinsen, Stijn H. P. Schoen, Cor D. Keijser, Bart J. F. Ossendrijver, Michel Hofstra, L. Marije Brouns, Stan J. J. Shinkai, Akeo van der Oost, John Staals, Raymond H. J. SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation |
title | SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation |
title_full | SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation |
title_fullStr | SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation |
title_full_unstemmed | SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation |
title_short | SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation |
title_sort | scope enables type iii crispr-cas diagnostics using flexible targeting and stringent carf ribonuclease activation |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8376896/ https://www.ncbi.nlm.nih.gov/pubmed/34413302 http://dx.doi.org/10.1038/s41467-021-25337-5 |
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