Fig. 4
From: Successful generation of epigenetic disease model mice by targeted demethylation of the epigenome
Generation of epigenome-edited mice by stable expression of epigenome editing factors in mice. a Schematics for generation of epigenome-edited mice. Linearized all-in-one vector (Fig. 1c) was introduced into the Rosa26 locus, which was cut by the CRISPR/Cpf1 system. b Epigenome-edited newborn mice were generated by introduction of a vector expressing gRNAs for both H19-DMR and H19-promoter (H19-DMR + P) or gRNAs for only H19-DMR. Control mice were generated by introduction of vector with TETCD mutant (TETCD-free) or vector without gRNAs (gRNA-free). Significant demethylation in H19-DMR and promoter regions was observed in the H19-DMR+P and H19-DMR integrated newborn mice. Interestingly, the mice in which only gRNAs for H19-DMR were introduced also showed demethylation in the H19-promoter region. c Expression analysis by qPCR. Vector-integrated newborn mice showed upregulation of H19 and downregulation of Igf2 expression. d Reduction of body weight was observed in newborn mice in which the vector with H19 gRNAs was integrated. n indicates the number of mice used for the analysis. Error bars, mean ± s.d. * P < 0.05, **P < 0.01, ***P < 0.001, n.s., not significant (two-tailed Student’s t test)