Identification of the 187 bp EphA7 Genomic DNA as the Dorsal Midline-Specific Enhancer of the Diencephalon and Mesencephalon

Abstract

EphA7 is a key molecule in regulating the development of the dien- and mesencephalon. To get insight into the mechanism of how EphA7 gene expression is regulated during the dorsal specification of the dien- and mesencephalon, we investigated the cis-acting regulatory sequence driving EphA7 to the dorsal midline of the dien- and mesencephalon. Transgenic LacZ reporter analysis, using overlapping EphA7 BACs, was used to narrow down the dorsal midline-specific enhancer, revealing the 25.3 kb genomic region as the enhancer candidate. Strikingly, this genomic DNA was located far downstream of the EphA7 transcription start site, +302.6 kb to +327.9 kb. Further enhancer mapping, using comparative genomic analysis and transgenic methods, showed that the 187 bp genomic DNA alone, approximately 305 kb downstream of the EphA7 transcription start site, was sufficient to act as the dorsal midline-specific enhancer of EphA7. Importantly, our results indicate that the 187 bp dorsal midline-specific enhancer is critically regulated by homeobox transcription factors during the development of the dien- and mesencephalon.

Tnni1b-ECR183-d2, an 87 bp cardiac enhancer of zebrafish

“…The previously reported gene-related enhancers are mostly hundreds to thousands of base pairs in length ( Abbasi et al, 2010 ; Anwar et al, 2015 ). Interestingly, researchers also found that some enhancers are small, such as a 187 bp dorsal midline-specific enhancer and a 44 bp vit-2 enhancer ( Charité et al, 2001 ; Kim, Park & Park, 2015 ; Goszczynski et al, 2016 ). These studies suggest that some distant and small enhancers are still capable of regulating gene expression.…”