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. 2017 Dec 14;46(2):873–885. doi: 10.1093/nar/gkx1264

Figure 3.

Figure 3.

Effect of T4 DNA modifications on type I-E CRISPR–Cas sgRNA mediated DNA targeting. (A) Schematic of DNA targeting and R-loop formation by Cascade. Modified cytosine residues are indicated in red. (B) Cleavage assay of Cas3 in conjunction with Cascade on 98 bp modified targets, indicated by black arrow. The marker is indicated by white arrows. Cascade effector complexes are loaded with either targeting crRNA (T crRNA) or non-targeting crRNA (NT crRNA). Restriction products of Cas3 are of undefined length. (C). Electrophoretic Mobility Shift Assay (EMSA) of Cascade on target DNA containing C, 5-hmC or 5-ghmC (indicated by black arrow) at increasing protein concentrations [nM]. Fraction of bound target is indicated by white arrows, dotted lines represent separate gels.