SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation
Abstract
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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Aug 19, 2021
- Source ID
- 10.1038/s41467-021-25337-5
Entities
People
- Akeo Shinkai
- Bart J. F. Keijser
- Cor D. Schoen
- David W Taylor
- Jack P. K. Bravo
- John van der Oost
- Jurre A. Steens
- L. Marije Hofstra
- Michel Ossendrijver
- Raymond H J Staals
- Stan J. J. Brouns
- Stijn H. P. Prinsen
- Yifan Zhu
Organizations
- Army Research Office
- Cancer Prevention and Research Institute of Texas
- Dutch Research Council
- Finnish Medical Society Duodecim
- National Institute of General Medical Sciences
- Netherlands Institute for Health Services Research
- Robert A. Welch Foundation
- Robert J Kleberg Jr and Helen C Kleberg Foundation