The Role of KDM2A and H3K36me2 Demethylation in Modulating MAPK Signaling during Neurodevelopment [ChIP-seq]
To investigate how KDM2A regulates downstream genes and affects NPC function and the role of KDM2A in histone methylation, we performed KDM2A and H3K36me2 ChIP-Seq. By analyzing both datasets together, we identified 2,051 overlapping peaks that were shared by both KDM2A and H3K36me2, The enriched genes were found to be associated with generating neurons, central nervous system development and cell proliferation. Furthermore, disease enrichment analysis revealed significant associations with ASD, ID and other neurodevelopmental disorders, indicating KDM2A's roles in neural development. Overall design: We generated KDM2A knockdown H9 ESC lines using shRNA lentivirus. We designed five siRNAs and selected the two most effective ones to create shRNA, which were then packaged into shRNA lentivirus (sh1, sh4). Next, we induced these cells into NPC. We performed KDM2A ChIP-Seq in control NPC and H3K36me2 ChIP-Seq in knockdown NPC (NC, sh1, sh4). ChIP-Seq analysis revealed that KDM2A preferentially binding to TSS regions of genes that related to neurogenesis and neuron differentiation. Furthermore, knockdown of KDM2A appeared to hinder H3K36me2 binding on regulatory elements of downstream genes.