Fig. 3

dCas9-Dnmt3a-mediated DNA methylation diminished APP expression in mouse brain. a Schematic of DNA methylation editing in the APP target region in the mouse brain in vivo. b Quantitative real-time PCR analysis of APP expression in the wild-type (WT) mouse brain infected with − 189 sgRNA and dCas9-Dnmt3a. Data are expressed as mean ± SEM (n = 3). **P < 0.01, two-sided Student’s t-test. c Western blot analysis of APP expression in the WT mouse brain transduced with − 189 sgRNA and dCas9-Dnmt3a. Data are expressed as mean ± SEM (n = 3). **P < 0.01, two-sided Student’s t-test. d Bisulfite sequencing analysis of the APP promoter region from the WT mouse brain and APP-KI mouse brain infected with − 189 sgRNA and dCas9-Dnmt3a. e Quantification of methylated amplicons. In 3 independent experiments, 10 to 30 sequencing data were measured in total. Data are expressed as mean ± SEM. **P < 0.01, two-sided Student’s t-test. f Quantitative real-time PCR analysis of predicted off-target genes in the WT mouse hippocampus transduced with − 189 sgRNA and dCas9-Dnmt3a. Data are expressed as mean ± SEM (n = 3). *P < 0.05, two-sided Student’s t-test. g Quantitative real-time PCR analysis of APP expression in the APP-KI mouse hippocampus transduced with − 189 sgRNA and dCas9-Dnmt3a. Data are expressed as mean ± SEM (n = 3). **P < 0.01, two-sided Student’s t-test. h Western blot analysis of APP in the APP-KI mouse hippocampus transduced with − 189 sgRNA and dCas9-Dnmt3a. Data are expressed as mean ± SEM (n = 3). **P < 0.01, two-sided Student’s t-test. The images in c, h are representative of three or more similar experiments