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. 2019 Jun 6;4(11):e128886. doi: 10.1172/jci.insight.128886

Figure 5. Site-specific methylation editing of the TNF DMR by dCas9-Tet1 increases TNF transcript level.

Figure 5

(A) Schematic representation of methylome editing of the TNF DMR by dCas9-Tet1 and specific sgRNAs. ChromHMM color code is in Figure 1C. (B) Methylation level of 13 individual CpG sites at the TNF DMR of human kidney tubules from control and DKD patients. (C) Upper panel: Methylation level of individual CpGs in the TNF DMR from dCas9-Tet1CD cells (HEK293) with or without DMR-targeting sgRNAs. Bottom panel: Bisulfite sequencing analysis of specific cells. (D) Relative transcript level (qRT-PCR) of TNF in control, dCas9-Tet1CD cells with DMR sgRNAs or NC sgRNA, and dCas9-Tet1CD_IM cells with DMR sgRNAs. (E) Relative transcript levels of genes associated with cell death in control, dCas9-Tet1CD or dCas9-Tet1CD_IM cells with DMR sgRNAs. All data are presented as means ± SEM of 3 experiments and analyzed by 1-way ANOVA with Tukey’s post hoc test.