A Ctnnb1 enhancer regulates neocortical neurogenesis by controlling the abundance of intermediate progenitors

beta-catenin-dependent canonical Wnt signaling plays a plethora of roles in neocortex (Ncx) development, but its function in regulating the abundance of intermediate progenitors (IPs) is elusive. Here we identified neCtnnb1, an evolutionarily conserved cis-regulatory element with typical enhancer features in developing Ncx. neCtnnb1 locates 55 kilobase upstream of and spatially close to the promoter of Ctnnb1, the gene encoding beta-catenin. CRISPR/Cas9-mediated activation or interference of the neCtnnb1 locus enhanced or inhibited transcription of Ctnnb1. neCtnnb1 drove transcription predominantly in the subventricular zone of developing Ncx. Knock-out of neCtnnb1 in mice resulted in compromised expression of Ctnnb1 and the Wnt reporter in developing Ncx. Importantly, knock-out of neCtnnb1 lead to reduced production and transit-amplification of IPs, which subsequently generated fewer upper-layer Ncx projection neurons (PNs). In contrast, enhancing the canonical Wnt signaling by stabilizing beta-catenin in neCtnnb1-active cells promoted the production of IPs and upper-layer Ncx PNs. ASH2L was identified as the key trans-acting factor that associates with neCtnnb1 and Ctnnb1's promoter to maintain Ctnnb1's transcription in both mouse and human Ncx progenitors. These findings advance understanding of transcriptional regulation of Ctnnb1, and provide insights into mechanisms underlying Ncx expansion during development.

Automated summary

This study identified the neCtnnb1 element, which plays a regulatory role in the development of the neocortex. neCtnnb1 drives the transcription of the Ctnnb1 gene and is predominantly active in the subventricular zone of the developing neocortex. Knocking out neCtnnb1 leads to reduced production and amplification of intermediate progenitors, resulting in fewer upper-layer neocortex projection neurons. Understanding the transcriptional regulation of Ctnnb1 contributes to our knowledge of the mechanisms underlying neocortex expansion during development.