SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation

SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation

Steens, Jurre A. (Wageningen University & Research; Scope Biosciences, Wageningen)
Zhu, Y. (Wageningen University & Research)
Taylor, David W. (University of Texas at Austin; Dell Medical School, Austin)
Bravo, Jack P.K. (University of Texas at Austin)
Prinsen, Stijn H.P. (Scope Biosciences, Wageningen)
Schoen, Cor D. (Wageningen University & Research)
Keijser, Bart J.F. (TNO)
Ossendrijver, Michel (TNO)
Brouns, S.J.J. (TU Delft BN/Stan Brouns Lab)

2021

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.

http://resolver.tudelft.nl/uuid:45777a1d-de7e-4bc0-b1a8-dacae7895b40

https://doi.org/10.1038/s41467-021-25337-5

2041-1723

Nature Communications, 12 (1)

Institutional Repository

journal article

© 2021 Jurre A. Steens, Y. Zhu, David W. Taylor, Jack P.K. Bravo, Stijn H.P. Prinsen, Cor D. Schoen, Bart J.F. Keijser, Michel Ossendrijver, S.J.J. Brouns, More Authors