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.

中文翻译：

---

斑马鱼 emx2 下游增强子 CNE2.04 的组织特异性相关区域和基序

背景

EMX2的表达水平在神经系统和癌症的发展中起着重要作用。CNE2.04 是emx2下游的保守增强子，以与emx2相似的模式驱动荧光蛋白表达。

方法

在斑马鱼卵的单细胞阶段，构建了 CNE2.04 截短或基序突变的转基因报告质粒，并用 Tol2 mRNA 注射到斑马鱼受精卵中。在 24、48 和 72 hpf 观察绿色荧光表达模式，并在 48 hpf 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，而CNE2.04-L400的组织特异性明显改变。基序CCCCTC突变明显改变了增强子的活性，而基序CCGCTC突变也改变了它。

结论

由于与组织特异性相关，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。