The tissue-specificity associated region and motif of an emx2 downstream enhancer CNE2.04 in zebrafish

IF 1 4区生物学 Q4 DEVELOPMENTAL BIOLOGY Gene Expression Patterns Pub Date : 2022-09-01 DOI:10.1016/j.gep.2022.119269

Xudong Chen, Qi Zhang, Jia Lin, Yinglan Zhang, Yawen Zhang, Yiting Gui, Ruizhi Zhang, Ting Liu, Qiang Li

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Abstract

Background

Expression level of EMX2 plays an important role in the development of nervous system and cancers. CNE2.04, a conserved enhancer downstream of emx2, drives fluorescent protein expression in the similar pattern of emx2.

Methods

CNE2.04 truncated or motif-mutated transgenic reporter plasmids were constructed and injected into the zebrafish fertilized egg with Tol2 mRNA at the unicellular stage of zebrafish eggs. The green fluorescence expression patterns were observed at 24, 48, and 72 hpf, and the fluorescence rates of different tissues were counted at 48 hpf.

Results

Compared to CNE2.04, CNE2.04-R400 had comparable enhancer activity, while the tissue specificity of CNE2.04-L400 was obviously changed. Motif CCCCTC mutation obviously changed the enhancer activity, while motif CCGCTC mutations also changed it.

Conclusion

Due to their correlation with tissue specificity, CNE2.04-R400 is associated with the tissue-specificity of CNE2.04, and motif CCCCTC plays an important role in the enhancer activity of CNE2.04.

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emx2下游增强子CNE2.04在斑马鱼中的组织特异性相关区域和基序

EMX2的表达水平在神经系统和肿瘤的发生发展中起重要作用。CNE2.04是emx2下游的保守增强子，其驱动荧光蛋白的表达模式与emx2相似。方法构建截断或基元突变的转基因报告质粒，在斑马鱼卵单细胞期将Tol2 mRNA注入受精卵中。在24、48、72 hpf下观察绿色荧光表达模式，在48 hpf下计数不同组织的荧光率。结果与CNE2.04相比，CNE2.04- r400具有相当的增强子活性，但CNE2.04- l400的组织特异性明显改变。Motif CCCCTC突变明显改变了增强子活性，而Motif CCGCTC突变也改变了增强子活性。结论CNE2.04- r400与CNE2.04的组织特异性相关，CCCCTC基序在CNE2.04的增强子活性中起重要作用。

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来源期刊

Gene Expression Patterns 生物-发育生物学

CiteScore

2.30

自引率

0.00%

发文量

审稿时长

35 days

期刊介绍： Gene Expression Patterns is devoted to the rapid publication of high quality studies of gene expression in development. Studies using cell culture are also suitable if clearly relevant to development, e.g., analysis of key regulatory genes or of gene sets in the maintenance or differentiation of stem cells. Key areas of interest include: -In-situ studies such as expression patterns of important or interesting genes at all levels, including transcription and protein expression -Temporal studies of large gene sets during development -Transgenic studies to study cell lineage in tissue formation

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