Transgenesis in Hydra to characterize gene function and visualize cell behavior
Transgenesis in Hydra to characterize gene function and visualize cell behavior
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DOI：
10.1038/s41596-019-0173-3
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发表时间：
2019-07-01
期刊：
影响因子：
14.8
通讯作者：
Bosch, Thomas C. G.
中科院分区：
文献类型：
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作者：
Klimovich, Alexander;Wittlieb, Joerg;Bosch, Thomas C. G.
淡水水螅是一种刺胞动物，在许多领域被用作模式生物，包括发育机制的起源和进化、衰老、共生和宿主-微生物相互作用的研究。本文介绍了一种利用胚胎显微注射建立转基因水螅稳定系的方法。该方案分为三个阶段，包括(i)设计和制备转基因构建体，（ii）将载体显微注射到水螅的早期胚胎中，以及（iii）选择和富集马赛克动物，以开发统一的转基因无性系。一个转基因构建体的制备需要2周，3个月内可获得转基因系。该方法允许本构性或诱导性的功能增益和功能损失方法，以及单个细胞的体内追踪。携带转基因细胞的水螅揭示了控制动物发育的祖先回路的功能特性。
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.