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- W2973307612 abstract "The multi-subunit type I CRISPR-Cas surveillance complex Cascade uses its crRNA to recognize dsDNA targets. Recognition involves DNA unwinding and base-pairing between the crRNA spacer region and a complementary DNA strand, resulting in formation of an R-loop structure. The modular Cascade architecture allows assembly of complexes containing crRNAs with altered spacer lengths that promise increased target specificity in emerging biotechnological applications. Here we produce type I-E Cascade complexes containing crRNAs with up to 57-nt-long spacers. We show that these complexes form R-loops corresponding to the designed target length, even for the longest spacers tested. Furthermore, the complexes can bind their targets with much higher affinity compared with the wild-type form. However, target recognition and the subsequent Cas3-mediated DNA cleavage do not require extended R-loops but already occur for wild-type-sized R-loops. These findings set important limits for specificity improvements of type I CRISPR-Cas systems." @default.
- W2973307612 created "2019-09-26" @default.
- W2973307612 creator A5001141726 @default.
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- W2973307612 date "2019-09-01" @default.
- W2973307612 modified "2023-10-12" @default.
- W2973307612 title "Decision-Making in Cascade Complexes Harboring crRNAs of Altered Length" @default.
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- W2973307612 doi "https://doi.org/10.1016/j.celrep.2019.08.033" @default.
- W2973307612 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6859484" @default.
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- W2973307612 hasPublicationYear "2019" @default.
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