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. 2020 Jun 4;48(12):6811–6823. doi: 10.1093/nar/gkaa477

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© The Author(s) 2020. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Unpaired bases at the 5′ end of gRNA slow the rate of DNA cleavage by the RuvC nuclease domain but not by the HNH nuclease domain. Quantified supercoiled plasmid cleavage assays from gels shown in Supplementary Figure S7B using Streptococcus thermophilus DGCC7710 CRISPR3 Cas9 WT (A), RuvC mutant D31A (B), or HNH mutant N891A (C). For clarity, only the disappearance of supercoiled (SC) DNA and appearance of linear (LIN) product for WT are compared to the appearance of nicked (OC) product for the nicking mutants. x-axes are split as 0–65 s and 75–300 s.