Identifying the binding sites for miRSome on miR-9 genomic locus with ChIP-seq

Recent studies have attributed multiple etiological factors for NDDs. miR-9/29a/107 is a key regulator in NDD. Therefore, understanding the upstream regulation of miR-9/29a/107 is critical for NDDs. We have used a unique multi-animal model-CC line that gives greater genetic diversity because miR upstream regulators could involve single or multiple factors. Using mirQTL mapping of CC we identified three candidates-Panx2/Polr1c/Mgea5. We showed that they modulate neuronal differentiation through miR-9. Next, we determined by ChIP that these genes are recruited to the regulatory region of the miR-9 locus. We showed that Panx2 is cleaved by RIP and translocated to the nucleus to interact with Polr1c and Mgea5. ChIP-combined-loop and 3C assays established the interaction on the miR-9 genomic locus through chromatin loops and histone modifications. We also proved that Panx2/Polr1c/Mgea5-miR-9 crosstalk to alter neuronal differentiation. Overall, we established a miRSome complex that govern transcriptional events during adult hippocampal neurogenesis. Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for Panx2, Polr1c and Mgea5 in duplicates with treatment and control files