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. 2019 Jun 21;2(3):172–185. doi: 10.1089/crispr.2019.0009

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© Nicholas C. Huston, et al. 2019; Published by Mary Ann Liebert, Inc.

This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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FIG. 6. — Cas9 molecules trapped in intermediate states may occlude target sites. In this model, a Cas9 RNP bound to substrate DNA is stuck in an inactive state (orange) and unable to cut a target successfully (i.e., low P_cut). Cas9 with higher P_cut has greater chance to cleave successfully once bound (blue). This decision is made heterogeneously throughout a population of bound RNPs, wherein each target sequence has a distribution of cleavage states (see Fig. 5). Potentially active Cas9 RNPs are unable to access and cleave the occluded site. Occluding Cas9 is maintained on substrate DNA by slow dissociation rates.