Kim, Jennifer. A novel enhancer of Dbx1 is able to drive neural tissue specific reporter gene expression. Retrieved from https://doi.org/doi:10.7282/T3JM29CP
DescriptionDbx1 is a homeobox gene that plays an essential role in the patterning of the central nervous system (CNS) during development. Its expression pattern is regionally restricted within the brain and spinal cord and has been linked to defining neuronal-specific domains. Enhancers, a major type of transcriptional regulatory element, are important in determining tissue specific gene expression. Here we have identified a novel enhancer of Dbx1 in order to understand mechanisms of spatial and temporal regulation of the Dbx1 gene. Using a multi-genome sequence alignment tool, a conserved non-coding region, Dbx1CR5 was identified as a putative enhancer of Dbx1. A green fluorescent protein (GFP) reporter assay system was used to verify the ability of Dbx1CR5 to drive neural tissue specific reporter gene expression in the developing chick CNS. GFP was detected between embryonic days 3 and 18 and was restricted to areas of the mesencephalon. Morphology and molecular staining of GFP+ tissues indicate that GFP is expressed in a heterogeneous population of cells. Within Dbx1CR5, multiple regions were identified to individually activate GFP expression in varying spatial patterns. Our findings support the notion that multiple binding elements reside within the 1135bp region of Dbx1CR5. Through this research we hope to identify factors that may one day be used to develop potential and novel therapeutics.