Fig. 3 | Nature Communications

Fig. 3

From: Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways

Fig. 3

Casilio-ME3 mediates dual delivery of TET1(CD) and NEIL2 to targeted genomic site. a Illustration of the indicated Casilio-ME platforms showing effector modules of PUFa fusion proteins used to transfect cells analyzed in panel (b). TET1(CD) (black), NEIL2 (blue), PUFa (light gray), occupancy of PBSa, amino (N) and carboxyl (C) termini of protein fusions are arbitrarily shown. b MLH1 mRNA relative levels (mean fold change ± S.E.M.; n = 3) in cells transfected with components of Casilio-ME1, Casilio-ME3.1, or Casilio-ME3.2 in the presence MLH1-sgRNAs or NT-sgRNA as indicated. Drawing of promoter regions with the MLH1-sgRNAs used (A-F), CpGs (lollipops), and TSS (arrow) is shown above the plot. ***p < 0.001, one-way ANOVA. c Schematic representation of the indicated Casilio-ME platforms showing effector modules of PUFa and PUFc protein fusions used to transfect cells analyzed in panel (d). TET1(CD) (black), NEIL2 (blue), PUFa (light gray), PUFc (orange), sgRNA containing both PBSa and PBSc, occupancy of PBSa and PBSc, amino (N) and carboxyl (C) termini of protein fusions are arbitrarily shown. d Plot showing MLH1 mRNA relative levels (mean fold change ± S.E.M.; n = 3) in cells transfected with components of Casilio-ME1, Casilio-ME3.3, or Casilio-ME3.4 in the presence of MLH1-sgRNAs. Drawing of promoter regions with the MLH1-sgRNAs used (A-F), CpGs (lollipops), and TSS (arrow) is shown above the plot. ***p < 0.005, one-way ANOVA

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