Fig. 1

H3K27me3 epigenome editing by dCas9-olEzh2 targeting hypomethylated promoters. a Schematic of dCas9, dCas9-olEzh2 and dCas9-olEzh2(∆SET) constructs and H3K27me3 induction caused by dCas9-olEzh2. b Schematic view of the dCas9-olEzh2 epigenome editing and injection experiments. sgRNA and mRNA were injected at the one-cell stage (stage 2). ChIP-qPCR was performed using the late blastula embryos (stage 11, 8 h after injection). RT-qPCR was performed using the pre-early gastrula embryos (stage 12, 10 h after injection), because ZGA occurs at the late blastula (stage 11) in medaka. c, g, k, n The epigenetic modification patterns around Arhgap35, Kita, Nanos3 and Dcx, sgRNAs (blue bars) and ChIP-qPCR product (black bars) positions. H3K27me3 (red) and H3K27ac (blue) ChIP-seq [27], DNase I-seq (black) [28] and DNA methylation [34] enrichment at the blastula stage are shown. d, e, h, i, l, m, o, p The results of ChIP-qPCR using anti-FLAG antibody (d, h, l, o) and anti-H3K27me3 antibody (e, i, l, m). H3K27me3 negative region (K27me3 NC) and H3K27me3 positive region (K27me3 PC) were used for ChIP control (described in Additional file 1: Fig. S2). f, j Arhgap35 and Pfkfb4a mRNA expression fold change. After expression levels were normalized to that of beta-actin, fold changes (sample/no injection) were calculated. Light blue, gray and orange bars in each bar graph represent no injection, sgRNAs/dCas9 injection and sgRNAs/dCas9-olEzh2 injection, respectively. (Tukey–Kramer test and only in Fig. 1f, j Student’s t test, *p < 0.1, **p < 0.05, ***p < 0.01, n = 3 biological replicates and only in Fig. 1f, j n = 6 biological replicates, error bars are s.d., p values of each comparison are shown only if the p value is under 0.1.)