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

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 6. — Effect of T4 DNA modifications on type V-A CRISPR–Cas sgRNA mediated DNA targeting. (A) Schematic of DNA targeting by Cas12a. Modified cytosine residues are indicated in red. Cleavage sites are indicated by black arrows. (B) Cleavage assay of Cas12a on 98 bp modified targets (indicated by black arrow). Cas12a is loaded with either targeting crRNA (C crRNA) or non-targeting crRNA (NC crRNA). Cleavage products of Cas12a are 49 and 44 bp. The marker is indicated by white arrows. (C) Electrophoretic Mobility Shift Assay (EMSA) of Cas12a 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.