CRISPR-Cas9-based editing of histone H3 arginine 17 reveals its methylation-dependent regulation of Yap signaling and early mouse embryo development (genomic DNA-seq data set). CRISPR-Cas9-based editing of histone H3 arginine 17 reveals its methylation-dependent regulation of Yap signaling and early mouse embryo development (genomic DNA-seq data set)

we use rAPOBEC-XTEN-Cas9n-UGI (BE3) to introduce a point mutation (R17H) into 8 loci of Hist1H3 by a single sgRNA and a pre-stop codon into Carm1 in early mouse embryo, and both strategies resulted in developmental defects in pre-implantation embryos and fetal stages with smaller or developmental-delayed embryos. Transcriptomic analysis revealed that Yap1 and cell cycle signaling pathways were dysregulated in Carm1 pre-stop and H3R17H embryos, and Yap1 overexpression could rescue the developmental defects. Our results establish the direct regulatory relationship between Carm1-mediated site-specific histone modifications and mouse embryo development, and we also demonstrate that Hippo-Yap1 acts downstream of Carm1-mediated H3R17me2a to regulate the embryonic development and size determination. Overall design: Examination of on-target and off-target induced by base editing

本研究采用rAPOBEC-XTEN-Cas9n-UGI（BE3）碱基编辑器，借助一条单向导RNA（single guide RNA, sgRNA）在小鼠早期胚胎的Hist1H3组蛋白的8个位点引入点突变R17H，并在精氨酸甲基转移酶1（CARM1）基因中引入提前终止密码子；两种编辑策略均使着床前胚胎及胎儿发育阶段出现发育缺陷，表现为胚胎体积偏小或发育迟缓。转录组分析结果显示，在CARM1提前终止突变体与H3R17H突变体胚胎中，Yes相关蛋白1（YAP1）及细胞周期信号通路均发生表达失调；且YAP1过表达可挽救上述发育缺陷。本研究明确了CARM1介导的位点特异性组蛋白修饰与小鼠胚胎发育之间的直接调控关联，同时证实Hippo-YAP信号通路位于CARM1介导的H3R17me2a（组蛋白H3精氨酸17位点二甲基化）下游，参与调控胚胎发育与体型大小确定。实验整体设计：检测碱基编辑诱导产生的靶向效应与脱靶效应。