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dc.contributor.authorLiao, Chunyu
dc.contributor.authorTtofali, Fani
dc.contributor.authorSlotkowski, Rebecca A
dc.contributor.authorDenny, Steven R
dc.contributor.authorCecil, Taylor D
dc.contributor.authorLeenay, Ryan T
dc.contributor.authorKeung, Albert J
dc.contributor.authorBeisel, Chase L
dc.date.accessioned2019-08-19T13:12:35Z
dc.date.available2019-08-19T13:12:35Z
dc.date.issued2019-07-03
dc.identifier.citationNat Commun. 2019 Jul 3;10(1):2948. doi: 10.1038/s41467-019-10747-3.en_US
dc.identifier.issn2041-1723
dc.identifier.pmid31270316
dc.identifier.doi10.1038/s41467-019-10747-3
dc.identifier.urihttp://hdl.handle.net/10033/621909
dc.description.abstractCRISPR-Cas systems inherently multiplex through CRISPR arrays—whether to defend against different invaders or mediate multi-target editing, regulation, imaging, or sensing. However, arrays remain difficult to generate due to their reoccurring repeat sequences. Here, we report a modular, one-pot scheme called CRATES to construct CRISPR arrays and array libraries. CRATES allows assembly of repeat-spacer subunits using defined assembly junctions within the trimmed portion of spacers. Using CRATES, we construct arrays for the single-effector nucleases Cas9, Cas12a, and Cas13a that mediated multiplexed DNA/RNA cleavage and gene regulation in cell-free systems, bacteria, and yeast. CRATES further allows the one-pot construction of array libraries and composite arrays utilized by multiple Cas nucleases. Finally, array characterization reveals processing of extraneous CRISPR RNAs from Cas12a terminal repeats and sequence- and context-dependent loss of RNA-directed nuclease activity via global RNA structure formation. CRATES thus can facilitate diverse multiplexing applications and help identify factors impacting crRNA biogenesis.en_US
dc.language.isoenen_US
dc.publisherSpringer-Natureen_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleModular one-pot assembly of CRISPR arrays enables library generation and reveals factors influencing crRNA biogenesis.en_US
dc.typeArticleen_US
dc.contributor.departmentHIRI, Helmholtz-Institut für RNA-basierte Infektionsforschung, Josef-Shneider Strasse 2, 97080 Würzburg, Germany.en_US
dc.identifier.journalNature Communicationsen_US
refterms.dateFOA2019-08-19T13:12:36Z
dc.source.journaltitleNature communications


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