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

IOS EDGE：实现弱电鱼的基因型-表型研究

• 批准号: 1644965
• 负责人: Jason Gallant
• 金额: $ 150万
• 依托单位: Michigan State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1644965&HistoricalAwards=false
• 关键词: IOS; EDGE; Enabling; Genotype; Phenotype

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.

自18世纪以来，电鱼一直是生物学中的模型系统，它提供了对生物电发生的本质、突触结构和复杂行为背后的大脑回路的深入了解。电鱼表现出许多有趣的特征或表型，但对这些表型如何在遗传物质或基因组中编码知之甚少。理解这种关系是21世纪生物学的一个广泛目标。 电鱼在试图理解基因组与其产生的表型之间的联系方面提供了一个独特的优势：两组电鱼在非洲和南美洲独立进化，提供了一个系统，所有类型的生物学家都可以通过内置复制来解决基因型-表型问题。 有越来越多的电鱼基因组数据;然而，没有功能性工具来研究这种联系。 该项目的目的是为电鱼模型系统开发一个强大的，可访问的，易于转移的遗传操作工具箱，以解决正在调查的所有问题，无论研究人员的背景如何。该项目的第一阶段是快速并行协议开发，该项目的第二阶段将是传播通过开发用于共享协议、数据和资源的基于网络的软件而获得的知识，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，为从事科学和技术职业做准备。拟议项目分为两个阶段。 该项目的第一阶段是快速并行方案开发，具体目标是：（1）使用CRISPR/Cas9技术产生稳定的突变体和转基因电鱼品系，（2）使用病毒介导的技术表达转基因材料，以及（3）使用吗啉代敲低特异性内源基因活性。 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，使社区能够靶向感兴趣的基因;（2）通过扩大该领域的参与和培训下一代电鱼生物学家来传播第一阶段制定的协议，以利用这些强大的新技术。 该项目大大加速了对NSF-IOS所有优先领域的基因型-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，以及广泛的表型数据，这些数据跨越了从分子到群体的生物学水平的分析。 这些研究的例子包括：（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）识别控制电感受器和电器官重复进化的发育机制，（5）光遗传学使能的脑回路解剖（6）将遗传变化与生殖隔离的进化联系起来。 这个项目的一个成果是使电鱼社区能够开发新的和令人兴奋的假设和项目，跨越生物学科的范围，并作为“大使”，以其他非规范的模型系统，应用新开发的技术。

项目成果

Sperm competition, sexual selection and the diverse reproductive biology of Osteoglossiformes

骨舌目的精子竞争、性选择和多样化的生殖生物学

• DOI: 10.1111/jfb.14779
• 发表时间: 2021
• 期刊: Journal of Fish Biology
• 影响因子: 2
• 作者: Koenig, Lauren A.;Gallant, Jason R.
• 通讯作者: Gallant, Jason R.

Silencing the Spark: CRISPR/Cas9 Genome Editing in Weakly Electric Fish

• DOI: 10.3791/60253
• 发表时间: 2019-10-01
• 期刊: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
• 影响因子: 1.2
• 作者: Constantinou, Savvas J.;Linh Nguyen;Gallant, Jason R.
• 通讯作者: Gallant, Jason R.