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.
