Evf2 enhancer long non-coding RNA regulation of gene expression in embryonic mouse brain

The Evf2 long non-coding RNA regulates genes adjacent and overlapping genes (Dlx5 and Dlx6), but it was not known if long range gene regulation occurs. In this study, we find that Evf2 regulates genes across a 27Mb region on mouse chromosome 6, and additional genes outside of mosue chromosome 6. We used microarrays to determine the effects of the Evf2 lncRNA on gene expression at sites of interenuron birth in a specific region of the mouse brain (ventral forebrain, MGE) at E13.5. Embryonic brain tissue from medial ganglionic eminence (MGE) at E13.5 were isolated from Evf2+/+ (wildtype) and Evf2TS/TS (lacking expression of the Evf2 lncRNA). Total RNA was isolated from pools of 4 MGE's per genotype. 5 pools were made from each genotype. Males and females were distinguished by Y-specific PCR analysis: Evf2+/+ n=2 males, n=3 females, Evf2TS/TS n=3 males, n=2 females