Meg8-DMR是在印迹Dlk1-Dio3结构域中发现的第一个母体甲基化DMR。Meg8-DMR的缺失增强了MLTC-1的迁移和侵袭,这取决于CTCF结合位点。然而,Meg8-DMR在小鼠发育过程中的生物学功能仍然未知。在这项研究中,使用CRISPR/Cas9系统在小鼠中产生434bp的Meg8-DMR基因组缺失。高通量和生物信息学分析显示,Meg8-DMR参与microRNA的调节:当缺失从母亲(Mat-KO)遗传时,microRNA的表达没有变化。然而,当从父亲(Pat-KO)和纯合(Homo-KO)发生缺失时,表达上调。然后,在WT与Pat-KO,Mat-KO,和Homo-KO,分别。随后,对这些DEGs进行了京都基因和基因组百科全书(KEGG)途径和基因本体论(GO)术语富集分析,以探索这些基因的功能作用。总的来说,测定502、128和165个DEG。GO分析表明,这些DEGs主要富集在Pat-KO和Home-KO的轴突发生中,而Mat-KO丰富了前脑发育。最后,IG-DMR的甲基化水平,Gtl2-DMR,Meg8-DMR,Dlk1、Gtl2和Rian的印迹状态没有受到影响。这些发现表明Meg8-DMR,作为二级监管区域,可以调节microRNAs的表达,同时不影响小鼠的正常胚胎发育。
Meg8-DMR is the first maternal methylated DMR to be discovered in the imprinted Dlk1-Dio3 domain. The deletion of Meg8-DMR enhances the migration and invasion of MLTC-1 depending on the CTCF binding sites. However, the biological function of Meg8-DMR during mouse development remains unknown. In this study, a CRISPR/Cas9 system was used to generate 434 bp genomic deletions of Meg8-DMR in mice. High-throughput and bioinformatics profiling revealed that Meg8-DMR is involved in the regulation of microRNA: when the deletion was inherited from the mother (Mat-KO), the expression of microRNA was unchanged. However, when the deletion occurred from the father (Pat-KO) and homozygous (Homo-KO), the expression was upregulated. Then, differentially expressed microRNAs (DEGs) were identified between WT with Pat-KO, Mat-KO, and Homo-KO, respectively. Subsequently, these DEGs were subjected to the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) term enrichment analysis to explore the functional roles of these genes. In total, 502, 128, and 165 DEGs were determined. GO analysis showed that these DEGs were mainly enriched in axonogenesis in Pat-KO and Home-KO, while forebrain development was enriched in Mat-KO. Finally, the methylation levels of IG-DMR, Gtl2-DMR, and Meg8-DMR, and the imprinting status of Dlk1, Gtl2, and Rian were not affected. These findings suggest that Meg8-DMR, as a secondary regulatory region, could regulate the expression of microRNAs while not affecting the normal embryonic development of mice.