Fig. 2: Targeted DNA Methylation of Mecp2 in vivo Using DNMT3L-DNMT3A-dCas9.
From: Locus-specific DNA methylation of Mecp2 promoter leads to autism-like phenotypes in mice
a Schematic illustration of experimental procedure for targeted DNA methylation in vivo. Constructs expressing DNMT3L-DNMT3A-dCas9 or DNMT3L-DNMT3Amut-dCas9 along with sgRNAs targeting the Mecp2 TSS region were pooled together and injected into mouse zygotes as indicated. b DNA methylation of targeted regions in the tail of male mice was analyzed by bisulfite sequencing. Each dot indicates one individual mouse. n = 8 mice/group; Data were statistically analyzed by Student’s t test (***p < 0.001), and shown as the mean ± s.e.m. c Representative methylation pattern in the tails of male mice as determined by bisulfite sequencing. d DNA methylation of target regions in the hippocampus (left) and parietal cortex (right) of male mice were analyzed by bisulfite sequencing. Each dot indicates one individual mouse. n = 8 mice/group; Data were statistically analyzed by Student’s t test (***p < 0.001), and shown as the mean ± s.e.m. e Methylation in the tail of male mice. Black portion of the circles indicates the methylation ratio in each CpG site. The results include all the sequencing data from mice described in (b). The Mecp2 sgRNAs (2–4) are indicated by blue rectangles.
