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. 2014 Nov 1;28(21):2432–2443. doi: 10.1101/gad.250712.114

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© 2014 Hale et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 4. — Requirements for target binding by the Cmr complex. (A) Cmr protein requirements for target RNA binding. The indicated recombinant Cmr proteins and unlabeled 45-nt crRNA were combined and incubated with 5′ end-labeled complementary target RNA. Complex formation was analyzed by native gel electrophoresis. (B) Target binding specificity. Recombinant Cmr1–6 and unlabeled 45-nt crRNA were combined and incubated with the indicated radiolabeled RNA or DNA substrates. Substrates are illustrated below each lane; asterisk indicates label location and substrate strand. “Target” substrates (gray) are complementary to the crRNA. “Nontarget” substrates (orange) are not complementary (equivalent to guide sequence of crRNA). Diagrams at the left of the panel indicate the electrophoretic positions of unbound labeled substrate (in the presence of crRNA) and crRNPs (in the presence of proteins).