New Tools for the Identification of Developmentally Regulated Enhancer Regions in Embryonic and Adult Zebrafish
New Tools for the Identification of Developmentally Regulated Enhancer Regions in Embryonic and Adult Zebrafish
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DOI：
10.1089/zeb.2012.0775
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发表时间：
2013-03-01
期刊：
影响因子：
2
通讯作者：
Harris, Matthew P.
中科院分区：
文献类型：
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作者：
Levesque, Mitchell P.;Krauss, Jana;Harris, Matthew P.
我们进行了筛选，以确定在斑马鱼中驱动组织特异性 Gal4 表达的发育调节增强子。我们获得了 63 个在胚胎或成年斑马鱼中具有表达模式的稳定转基因系。使用来自青鳉edar基因座的新鉴定的最小启动子产生了一组相对公正的表达模式，代表源自所有胚层的许多组织类型。随后对所选细胞系的详细表征表明，Gal4 驱动的 GFP 在多种组织中均具有强烈且可重复的表达，包括中枢和周围神经系统的神经元、色素细胞、红细胞和皮周细胞。通过筛选成年斑马鱼的 GFP 表达，我们还分离了在成年斑马鱼组织中表达的品系，包括鳞片、鳍条和关节。我们确定的新型高效最小启动子和大量反式激活驱动系将为斑马鱼群体提供有用的资源，用于进一步增强子陷阱筛选以及体内组织特异性过程的精确研究。
We have conducted a screen to identify developmentally regulated enhancers that drive tissue-specific Gal4 expression in zebrafish. We obtained 63 stable transgenic lines with expression patterns in embryonic or adult zebrafish. The use of a newly identified minimal promoter from the medaka edar locus resulted in a relatively unbiased set of expression patterns representing many tissue types derived from all germ layers. Subsequent detailed characterization of selected lines showed strong and reproducible Gal4-driven GFP expression in diverse tissues, including neurons from the central and peripheral nervous systems, pigment cells, erythrocytes, and peridermal cells. By screening adults for GFP expression, we also isolated lines expressed in tissues of the adult zebrafish, including scales, fin rays, and joints. The new and efficient minimal promoter and large number of transactivating driver-lines we identified will provide the zebrafish community with a useful resource for further enhancer trap screening, as well as precise investigation of tissue-specific processes in vivo.