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. 2017 Nov 23;7:16174. doi: 10.1038/s41598-017-16279-4

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Evaluation of Cas9 orthologues in a PAM-specific system targeted to prevalent TGFBI mutations Guide RNA tested shown in purple, PAM utilised shown in green and mutation shown in red. (a) 22 and 21 nucleotide guides were designed to target the novel S.aureus Cas9 PAM generated by R555W. Both guide lengths were targeted to a luciferase reporter plasmid containing either a wild-type or mutant TGFBI sequence to determine potency and allele specificity. (b) A guide utilising the novel mutant AsCpf1 PAM generated by R124L was targeted to a luciferase reporter plasmid containing either a wild-type or mutant TGFBI sequence to determine potency and allele specificity.