比较序列分析和功能验证确定了围绕 zic5 和 zic2a 的视网膜特异性增强子。

Comparative sequence analysis and functional validation identified a retina-specific enhancer around zic5 and zic2a.

机构信息

Translational Medical Center for Development and Disease, Institute of Pediatrics, Shanghai Key Laboratory of Birth Defect, Children's Hospital of Fudan University, Shanghai, China.

出版信息

Gene Expr Patterns. 2021 Mar;39:119162. doi: 10.1016/j.gep.2020.119162. Epub 2020 Dec 28.

DOI:10.1016/j.gep.2020.119162

PMID:33359848

Abstract

The spatiotemporal development of vertebrate retina is regulated by a variety of genes. The zinc finger transcription factors zic5 and zic2a are located close to each other in the chromosome. They have similar expression patterns, and both play important roles in the development of the retina and nervous system. Here, we used ECR browser and gfp fluorescence report experiment to identify a 290bp enhancer sequence ECR3, which is located at 3 kb upstream of zic5 and 10 kb downstream of zic2a, and it can drive the specific expression of gfp in the retina. pT2KXIGQ-ECR3 was used to construct a transgenic zebrafish line Tg(ECR3-290: gfp) which first exhibits specific green fluorescence in the whole retina area at 24hpf. Then the expression region was gradually limited to ganglion cell layer (GCL) and lasted through adulthood. This expression pattern is highly consistent with the zic5 and zic2a at retina. These results indicate that the 290bp enhancer might be an important element to regulate the expression of zic5 and zic2a genes in ganglion cells, and this transgenic line is an important tool for studying the development of retina.

摘要

脊椎动物视网膜的时空发育受多种基因调控。锌指转录因子 zic5 和 zic2a 位于染色体上彼此靠近的位置。它们具有相似的表达模式，在视网膜和神经系统的发育中都发挥着重要作用。在这里，我们使用 ECR 浏览器和 GFP 荧光报告实验鉴定了一个 290bp 的增强子序列 ECR3，它位于 zic5 的上游 3kb 和 zic2a 的下游 10kb，能够驱动 GFP 在视网膜中的特异性表达。pT2KXIGQ-ECR3 被用来构建一个转基因斑马鱼系 Tg(ECR3-290: gfp)，它在 24hpf 时首先在整个视网膜区域表现出特异性的绿色荧光，然后表达区域逐渐局限于神经节细胞层（GCL）并持续到成年。这种表达模式与视网膜中的 zic5 和 zic2a 高度一致。这些结果表明，290bp 的增强子可能是调节神经节细胞中 zic5 和 zic2a 基因表达的一个重要元件，该转基因系是研究视网膜发育的重要工具。