Fig. 3
From: Successful generation of epigenetic disease model mice by targeted demethylation of the epigenome
Generation of epigenome-edited mice by transient expression of epigenome editing factors in preimplantation embryos. a Schematic for generation of epigenome-edited mice. In vitro transcribed mRNA was introduced into zygotes. b Epigenome editing factors (GFP) were expressed in almost all embryos 1 day after introduction. Scale bars, 200 μm. c Percentages of DNA methylation by COBRA in H19-DMR (m1-m4) and promoter (p1-p4) in epigenome-edited (epiedit) blastocyst (gRNAs for both H19-DMR and H19-promoter) and various control blastocysts. Non-target control was generated by introduction of gRNA for RASSF1A (human specific sequence). Blastocyst sample introduced all components shows significant demethylation in H19-DMR compared to other control sample. d Epigenome-edited newborn mice were generated by introduction of gRNAs for both H19-DMR and H19-promoter (H19-DMR + P, red dot) or gRNAs for only H19-DMR (yellow dot). Most newborn mice did not show a change in methylation, whereas four mice (arrows) showed apparent demethylation compared with control mice. e Expression analysis by qPCR. The four demethylated newborn mice showed downregulation of Igf2 expression compared with methylated control mice. f The four demethylated newborn mice showed significantly lower body weight than methylated control mice. Error bars, mean ± s.d. *P < 0.05, **P < 0.01, n.s., not significant (two-tailed Student’s t test)