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. 2021 Jun 16;49(15):8785–8795. doi: 10.1093/nar/gkab507

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© The Author(s) 2021. 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 2. — Comparison of SpCas9, Cas9-NG, xCas9, SpG and SpRY at NGH loci. (A) Editing efficiency for SpCas9 and indicated PAM-flexible variants at 15 NGH (NGA, NGC or NGT) loci in HEK293T cells detected by PEM-seq. (B) Off-target numbers at indicated NGH loci detected by PEM-seq. ‘-’ indicates nearly no editing activity. * indicates low editing efficiency for all variants. PAM and locus information are marked above. (C) Venn diagram for off-targets of Cas9-NG, SpG and SpRY at EMX1-AGC locus detected by PEM-seq. (D) Consensus analysis by weblogo for PAM sequence for Cas9-NG, SpG and SpRY off-targets for all NGH loci detected by PEM-seq. (E) Statistics of sgRNA mismatch numbers between off-target and on target for all 15 NGH loci for indicated SpCas9 variants detected by PEM-seq.