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

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.