Zoological Institute, University of Kiel, Kiel, Germany.
Nat Protoc. 2019 Jul;14(7):2069-2090. doi: 10.1038/s41596-019-0173-3. Epub 2019 Jun 3.
The freshwater polyp Hydra is a cnidarian used as a model organism in a number of fields, including the study of the origin and evolution of developmental mechanisms, aging, symbiosis and host-microbe interactions. Here, we describe a procedure for the establishment of stable transgenic Hydra lines by embryo microinjection. The three-stage protocol comprises (i) the design and preparation of a transgenic construct, (ii) the microinjection of the vector into early embryos of Hydra vulgaris, and (iii) the selection and enrichment of mosaic animals in order to develop uniformly transgenic clonal lines. The preparation of a transgenic construct requires ~2 weeks, and transgenic lines can be obtained within 3 months. The method allows constitutive or inducible gain- and loss-of-function approaches, as well as in vivo tracing of individual cells. Hydra polyps carrying transgenic cells reveal functional properties of the ancestral circuitry controlling animal development.
淡水水螅是一种刺胞动物,被广泛用作多个领域的模式生物,包括发育机制的起源和演化、衰老、共生和宿主-微生物相互作用的研究。在这里,我们描述了一种通过胚胎显微注射建立稳定的转基因水螅系的方法。该三阶段方案包括:(i)转基因构建体的设计和准备,(ii)将载体显微注射到普通水螅的早期胚胎中,以及(iii)选择和富集嵌合体动物,以开发均匀的转基因克隆系。转基因构建体的制备大约需要 2 周时间,而转基因系可以在 3 个月内获得。该方法允许进行组成型或诱导型的功能获得和功能丧失方法,以及对单个细胞的体内示踪。携带转基因细胞的水螅水螅揭示了控制动物发育的原始电路的功能特性。