IOS EDGE: Enabling Genotype-Phenotype Studies in Weakly Electric Fish

自18世纪以来，电鱼一直是生物学中的一个模型系统，它提供了对生物电发生的本质、突触的结构以及复杂行为背后的大脑电路的洞察。电鱼表现出许多有趣的特征或表型，但人们对这些表型是如何在遗传物质或基因组中编码的知之甚少。理解这种关系是21世纪生物学的一个宏伟目标。在试图了解基因组和其产生的表型之间的联系方面，电鱼提供了一个独特的优势：在非洲和南美洲独立进化的两组电鱼，提供了一个系统，在这个系统中，各种类型的生物学家可以通过内置复制来解决基因表型问题。有越来越多的电鱼基因组数据；然而，还没有功能工具来研究这种联系。该项目的目的是开发一个强大的，可访问的，和易于转移的遗传操作工具箱，用于电动鱼类模型系统的所有正在调查的问题，而不考虑研究人员的背景。该项目的第一阶段是快速并行方案开发，第二阶段将是传播通过开发基于网络的软件以共享方案、数据和资源而获得的知识，并培训下一代电鱼生物学家--来自不同背景的本科生和研究生--使用这些技术为在科学和技术领域的职业生涯做准备。拟议的项目分为两个阶段。该项目的第一阶段是快速平行方案开发，具体目标是：(1)使用CRISPR/Cas9技术培育稳定的电鱼突变和转基因品系，(2)使用病毒介导法表达转基因材料，(3)使用吗啉抑制特定的内源基因活性。该项目的第二阶段是传播知识和改善社区基础设施，具体目标是：(1)建立和分发基因组资源和工具，使社区能够针对感兴趣的基因；(2)通过扩大该领域的参与和培训下一代电鱼生物学家来利用这些强大的新技术，传播在第一阶段制定的协议。该项目大大加快了对NSF-iOS所有优先领域中的基因-表型相互作用的研究，因为它结合了基因组操作的力量和全球可访问的模型系统，以及跨越从分子到种群的生物分析水平的广泛的表型数据。这类研究的例子包括：(1)深入了解离子通道功能(2)识别电感觉系统中的转导分子，(3)识别支配电感受器和电子器官重复进化的发育机制，(5)利用光遗传学对大脑回路进行解剖(6)将遗传变化与生殖隔离的进化联系起来。该项目的一个成果是使电鱼社区能够开发跨越生物学学科范围的新的和令人兴奋的假设和项目，并充当其他非规范模型系统的“大使”，以应用新开发的技术。

Electric fish have served as a model system in biology since the 18th century, providing insight into the nature of bioelectrogenesis, the structure of the synapse, and brain circuitry underlying complex behavior. Electric fishes exhibit many interesting characteristics or phenotypes, but very little is known about how these phenotypes are encoded in the genetic material or genome. Understanding this relation is a broad goal in 21st century biology. Electric fish offer a singular advantage in trying to understand the link between the genome and the phenotype it produces: two groups of electric fish evolved independently in Africa and South America, providing a system where biologists of all stripes can attack genotype-phenotype questions with built-in replication. There is a growing amount of electric fish genomic data; however, there are no functional tools to investigate this connection. The purpose of this project is to develop a robust, accessible, and easily transferable genetic manipulation toolbox for the electric fish model system full range of questions under investigation, regardless of a researchers' background. The first phase of the project is rapid parallel protocol development, and the second phase of the project will be to disseminate knowledge gained by developing web-based software for sharing protocols, data, and resources, and to train the next generation of electric fish biologists--undergraduate and graduate students from diverse backgrounds--in the use of these techniques in preparation for careers in science and technology. The proposed project, is divided into two phases. The first phase of the project is rapid parallel protocol development, with the specific aims to: (1) generate stable mutant and transgenic lines of electric fish using CRISPR/Cas9 technologies, (2) express transgenic material using viral-mediated techniques, and (3) knockdown specific endogenous gene activity using morpholinos. The second phase of the project is dissemination of knowledge and community infrastructure improvement, with the specific aims to: (1) establish and distribute genomic resources and tools that enable the community to target genes of interest and (2) disseminate protocols developed in Phase 1 by broadening participation in the field and training the next generation of electric fish biologists to harness these powerful new techniques. The project accelerates significantly research on genotype-phenotype interaction across the full range of NSF-IOS priority areas because it combines the power of genomic manipulation and a globally accessible model system with extensive phenotypic data that span biological levels of analysis from molecules to populations. Examples of such studies include: (1) insight into ion channel function (2) identification of transduction molecules in electrosensory systems, (3) identification of developmental mechanisms governing the repeated evolution of electroreceptors and electric organs, (5) optogenetics-enabled dissections of brain circuits (6) link genetic changes and the evolution of reproductive isolation. An outcome of this project is the enablement of the electric fish community to develop new and exciting hypotheses and projects spanning the gamut of biological disciplines, and act as "ambassadors" to other non-canonical model systems to apply newly developed techniques.