Regulation of chromatin accessibility and Zic binding at enhancers in the developing cerebellum

To identify chromatin mechanisms of neuronal differentiation, we characterized chromatin accessibility and gene expression in cerebellar granule neurons (CGNs) of the developing mouse. We used DNase-seq to map accessibility of cis-regulatory elements and RNA-seq to profile transcript abundance across postnatal stages of neuronal differentiation in vivo and in culture. We observed thousands of chromatin accessibility changes as CGNs differentiated and verified by H3K27ac ChIP-seq, reporter gene assays, and CRISPR-mediated activation that many of these regions function as neuronal enhancers. Motif discovery within differentially accessible chromatin regions suggested a novel role for the Zic family of transcription factors in CGN maturation. We confirmed the association of Zic with these elements by ChIP-seq, and demonstrated by knockdown that Zic1/2 are required to coordinate mature neuronal gene expression patterns. Together these data reveal chromatin dynamics at thousands of gene regulatory elements that facilitate gene expression patterns necessary for neuronal differentiation and function.