Stable transgenesis in Astyanax mexicanus using the Tol2 transposase system
Stable transgenesis in Astyanax mexicanus using the Tol2 transposase system
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DOI：
10.1002/dvdy.32
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发表时间：
2019-01
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通讯作者：
B. A. Stahl;R. Peuß;Brittnee McDole;A. Kenzior;James B. Jaggard;K. Gaudenz;Jaya Krishnan;S. McGaugh;Erik R. Duboue;A. Keene;Nicolas Rohner
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作者：
B. A. Stahl;R. Peuß;Brittnee McDole;A. Kenzior;James B. Jaggard;K. Gaudenz;Jaya Krishnan;S. McGaugh;Erik R. Duboue;A. Keene;Nicolas Rohner
墨西哥Astyanax mexicanus是一种成熟且广泛应用于进化和发育生物学研究的鱼类模型系统。这些鱼以水面形式存在，栖息在河流和30种不同的洞穴鱼种群中。由于越来越多的基因组方法可以识别基因型-表型关联，墨西哥拟南蝽作为一种新兴的模式生物的建立加快了对发育和行为进化基础的理解。尽管在新技术的应用方面取得了重要进展，但由于缺乏遗传模型系统中常用的转基因系，对墨西哥拟南芥进化和发展的深入机制认识受到限制。在这里，我们扩展了转基因的工具箱，通过描述两个新的稳定的转基因系，这些系是用高效的Tol2系统产生的，通常用于产生转基因斑马鱼。一个由斑马鱼泛素启动子与eGFP融合组成的稳定转基因系在Astyanax水面种群的整个发育过程中普遍表达eGFP。为了确定特定的细胞类型，我们给鱼注射了Cntnap2-mCherry构建物，该构建物标记了斑马鱼的侧线机械感觉神经元。引人注目的是，这两种结构似乎都标记了预测的细胞类型，这表明斑马鱼中的许多遗传工具和定义的启动子区域可以直接转移到洞穴鱼中。这些品系为Tol2转基因技术在墨西哥稻上的应用提供了原理证明。这些初始转基因品系的扩展将提供一个平台，以解决在寻求弥合基因型与表型差距的广泛重要问题。
Astyanax mexicanus is a well-established and widely used fish model system for evolutionary and developmental biology research. These fish exist as surface forms that inhabit rivers and 30 different populations of cavefish. The establishment of A. mexicanus as an emergent model organism for understanding the evolutionary basis of development and behavior has been accelerated by an increasing availability of genomic approaches to identify genotype-phenotype associations. Despite important progress in the deployment of new technologies, deep mechanistic insights into A. mexicanus evolution and development have been limited by a lack of transgenic lines commonly used in genetic model systems. Here, we expand the toolkit of transgenesis by characterizing two novel stable transgenic lines that were generated using the highly efficient Tol2 system, commonly used to generate transgenic zebrafish. A stable transgenic line consisting of the zebrafish ubiquitin promoter fused to eGFP expressed eGFP ubiquitously throughout development in a surface population of Astyanax. To define specific cell-types, we injected fish with a Cntnap2-mCherry construct that labels lateral line mechanosensory neurons in zebrafish. Strikingly, both constructs appear to label the predicted cell types, suggesting many genetic tools and defined promoter regions in zebrafish are directly transferrable to cavefish. The lines provide proof-of-principle for the application of Tol2 transgenic technology in A. mexicanus. Expansion on these initial transgenic lines will provide a platform to address broadly important problems in the quest to bridge the genotype to phenotype gap.