Suster Maximiliano L, Kikuta Hiroshi, Urasaki Akihiro, Asakawa Kazuhide, Kawakami Koichi
Division of Molecular and Developmental Genetics, National Institute of Genetics, Mishima, Shizuoka, Japan.
Methods Mol Biol. 2009;561:41-63. doi: 10.1007/978-1-60327-019-9_3.
The zebrafish (Danio rerio) is a useful model for genetic studies of vertebrate development. Its embryos are transparent and develop rapidly outside the mother, making it feasible to visualize and manipulate specific cell types in the living animal. Zebrafish is well suited for transgenic manipulation since it is relatively easy to collect large numbers of embryos from adult fish. Several approaches have been developed for introducing transgenes into the zebrafish germline, from the injection of naked DNA to transposon-mediated integration. In particular, the Tol2 transposable element has been shown to create insertions in the zebrafish genome very efficiently. By using Tol2, gene trap and enhancer trap vectors containing the GFP reporter gene or yeast transcription activator Gal4 gene have been developed. Here we outline methodology for creating transgenic zebrafish using Tol2 vectors, and their applications to visualization and manipulation of specific tissues or cells in vivo and for functional studies of vertebrate neural circuits.
斑马鱼（Danio rerio）是脊椎动物发育遗传学研究的有用模型。其胚胎是透明的，且在母体外发育迅速，这使得在活体动物中可视化和操纵特定细胞类型成为可能。斑马鱼非常适合转基因操作，因为从成年鱼中收集大量胚胎相对容易。已经开发了几种将转基因引入斑马鱼种系的方法，从注射裸DNA到转座子介导的整合。特别是，Tol2转座元件已被证明能非常有效地在斑马鱼基因组中产生插入。通过使用Tol2，已开发出含有绿色荧光蛋白（GFP）报告基因或酵母转录激活因子Gal4基因的基因捕获和增强子捕获载体。在这里，我们概述了使用Tol2载体创建转基因斑马鱼的方法，以及它们在体内可视化和操纵特定组织或细胞以及脊椎动物神经回路功能研究中的应用。
