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The functional verification and analysis of Fugu promoter of cardiac gene tnni1a in zebrafish

斑马鱼 河豚 生物 遗传学 基因 保守序列 发起人 基因亚型 调节顺序 吗啉
作者
Yiting Gui,Yawen Zhang,Qi Zhang,Xudong Chen,Feng Wang,Fang Wu,Yonghao Gui,Qiang Li
出处
期刊:Cells and development [Elsevier]
卷期号:171: 203801-203801
标识
DOI:10.1016/j.cdev.2022.203801
摘要

Troponin I type 1b ( Tnni1b ) is thought to be a novel isoform that is expressed only in the zebrafish heart. Knocking down of tnni1b can lead to cardiac defects in zebrafish. Although both the zebrafish tnni1b and human troponin I1 ( TNNI1 ) genes are thought to be closely associated with fatal cardiac development, the regulatory molecular mechanisms of these genes are poorly understood. Analyzing the functionally conserved sequence, especially in the noncoding regulatory region involved in gene expression, clarified these mechanisms. In this study, we isolated a 3 kb fragment upstream of Fugu tnni1a that can regulate green fluorescence protein (GFP) expression in a heart-specific manner, similar to the pattern of zebrafish homologue expression. Three evolutionarily conserved regions (ECRs) in the 5′-flanking sequence of Fugu tnni1a were identified by sequence alignment. Deletion analysis led to the identification of ECR2 as a core sequence that affects the heart-specific expression function of the Fugu tnni1a promoter. Interestingly, both the Fugu tnni1a promoter and ECR2 sequence were functionally conserved in zebrafish, although they shared no sequence similarity. Together, the findings of our study provided further evidence for the important role of tnni1a homologous in cardiac development and demonstrated that two functionally conserved sequences in the zebrafish and Fugu genomes may be ECRs, despite their lack of similarity. • A functional promoter of Fugu tnni1a was identified in zebrafish. • This identified Fugu tnni1a promoter is heart-specific and relatively short. • The promoter drives GFP expression in the same pattern as zebrafish tnni1b. • Three ECRs were identified in the promoter region through comparative studies. • Deletion of ECR2 results in loss of cardiac specificity of the promoter.
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