Effects of deletion Rack7 on gene expression in primary astrocyte during mouse postnatal development [RNA-Seq]

Dysregulation of epigenetic mechanisms play crucial roles in brain development and disease. Emerging evidence suggests that RACK7, an epigenetic reader protein, may play a role in brain development, but in vivo exploration of RACK7 in neural development and the underlying mechanisms are still lacking. Here, we established a conditional knock-out Rack7 mouse model and show that Rack7-deficient mice exhibit overt developmental defects. Multicomplex immunofluorescence staining (mIF) of cell-type specific markers indicates that the developmental defects are associated with compromised astrocyte development. Mechanistically, we surprisingly found RACK7 interacts with PRC2 complex, which acts as the histone H3K27 methyltransferase. Our results suggest RACK7 binding with PRC2 complex helps to establish the proper genomic location of histone H3K27me3 modification. Deletion of Rack7 in astrocyte leads to a remarkable decrease of H3K27me3 chromatin enrichment. genome-wide?. Functionally, we found RACK7 works together with H3K27me3 to prevent overactivation of Wnt signaling pathway and other astrocyte differentiation genes. Collectively, our study provides new insights into the cellular and molecular mechanisms underlying brain development regulated by RACK7. Overall design: In order to determine the impact of Rack7 loss on the transcriptome during astrocyte development, we extracted and cultured primary astrocyte from WT and Rack7-cKO mice brain at three time points, including neonatal (P1), P6 and P8, and performed RNA-seq analysis.