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. 2019 Aug 14;47(18):9637–9657. doi: 10.1093/nar/gkz709

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© The Author(s) 2019. 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 8. — Analysis of whole-genome methylation following epigenetic modulation of the IL6ST/MGAT3 promoters using DNMT3A-dSpCas9 and TET1-dSpCas9 fusion constructs. (A) Fraction of differentially methylated probes compared to mock controls against total filtered probes analyzed on the Infinium MethylationEPIC array. Labels DNMT3A-PRI/TET1-PRI and DNMT3A-SEC/TET1-SEC indicate ‘primary’ and ‘secondary cassette’, respectively. (B) Fraction of differentially methylated probes, which were hypermethylated or hypomethylated, compared to mock controls in cells transfected with DNMT3A-dSpCas9 and TET1-dSpCas9 fusions. (C and E) Dots represent CpG sites targeted by DNMT3A-dSpCas9 and TET1-dSpCas9 fusion constructs. Lines show pattern of methylation (C) and demethylation (E) change at CpG island of IL6ST and MGAT3 genes. Comparison of CpG methylation patterns induced by DNMT3A-dSpCas9 expressed under strong (‘primary cassette’) or weak (‘secondary cassette’) promoter on probes located 2500 bp up- and downstream of the targeted region (shaded in gray) of IL6ST is shown on (C). Changes to β-values in the two probes lying within this region are illustrated on (D). (E) Comparison of DNA methylation patterns induced by TET1-dSpCas9 primary and secondary cassettes 2500 bp up- and downstream of the targeted region of the MGAT3 promoter region. The probe cg21461856 is the only one lying within this region (indicated with the arrow). Changes to β-values in cg21461856 are illustrated in the graph (F). (G) Venn diagrams illustrating overlap between DMPs induced by the primary and secondary cassettes of DNMT3A-dSpCas9 and TET1-dSpCas9 fusions. * P < 0.05.