Gehrig Jochen, Reischl Markus, Kalmár Eva, Ferg Marco, Hadzhiev Yavor, Zaucker Andreas, Song Chengyi, Schindler Simone, Liebel Urban, Müller Ferenc
Institute of Toxicology and Genetics, Forschungszentrum Karlsruhe, Eggenstein-Leopoldshafen, Germany.
Nat Methods. 2009 Dec;6(12):911-6. doi: 10.1038/nmeth.1396. Epub 2009 Nov 8.
Zebrafish embryos offer a unique combination of high-throughput capabilities and the complexity of the vertebrate animal for a variety of phenotypic screening applications. However, there is a need for automation of imaging technologies to exploit the potential of the transparent embryo. Here we report a high-throughput pipeline for registering domain-specific reporter expression in zebrafish embryos with the aim of mapping the interactions between cis-regulatory modules and core promoters. Automated microscopy coupled with custom-built embryo detection and segmentation software allowed the spatial registration of reporter activity for 202 enhancer-promoter combinations, based on images of thousands of embryos. The diversity of promoter-enhancer interaction specificities underscores the importance of the core promoter sequence in cis-regulatory interactions and provides a promoter resource for transgenic reporter studies. The technology described here is also suitable for the spatial analysis of fluorescence readouts in genetic, pharmaceutical or toxicological screens.
斑马鱼胚胎为各种表型筛选应用提供了高通量能力与脊椎动物复杂性的独特结合。然而，需要成像技术自动化以挖掘透明胚胎的潜力。在此，我们报告了一种高通量流程，用于在斑马鱼胚胎中记录特定结构域的报告基因表达，目的是绘制顺式调控模块与核心启动子之间的相互作用。自动化显微镜与定制的胚胎检测和分割软件相结合，基于数千个胚胎的图像，实现了202种增强子 - 启动子组合的报告基因活性的空间配准。启动子 - 增强子相互作用特异性的多样性强调了核心启动子序列在顺式调控相互作用中的重要性，并为转基因报告基因研究提供了启动子资源。这里描述的技术也适用于遗传、药物或毒理学筛选中荧光读数的空间分析。
