Tnni1b-ECR183-d2, an 87 bp cardiac enhancer of zebrafish
Abstract
Several heart malformations are associated with mutations in the regulatory regions of cardiac genes. Troponin I type 1b (tnni1b) is important for the formation of the atrioventricular canal in zebrafish hearts; however, the regulation of tnni1b is poorly understand. We aimed to identify a small but functional enhancer that is distal to tnni1b. Evolutionary Conserved Region (ECR) Browser was used to analyze the 219 kb zebrafish and human genomes covering the tnni1b gene as well as the 100 kb regions upstream and downstream of tnni1b. Putative transcription factor binding sites (TFBSs) were analyzed using JASPAR and PROMO, and the enhancer activity was identified using zebrafish embryos and the luciferase reporter assay. A correlation analysis between the enhancer and transcription factors (TFs) was performed via TF overexpression and TFBS mutation experiments and the electrophoretic mobility shift assay (EMSA). To analyze the conservation between zebrafish and human enhancers, human DNA fragments were functionally verified. Images were captured and analyzed by fluorescence microscopy or confocal microscopy. Combined with comparative analysis and functional validation, we identified a 183 bp ECR (termed tnni1b-ECR183) that was l...Continue Reading
References
Characterization and regulation of the hb9/mnx1 beta-cell progenitor specific enhancer in zebrafish.
Development of enhancer-trapping and -detection vectors mediated by the Tol2 transposon in zebrafish
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