A New Opportunity for Protecting Zebrafish Genetic Resources: Development of a Reproducible Repository System for Community-driven Cryopreservation

University Of Oregon, Eugene OR

Investigators

Abstract

PROJECT SUMMARY/ABSTRACT: Zebrafish (Danio rerio) is one of the most used animal models in biomedical research, and laboratories around the world produce thousands of new genetic lines each year. Establishment of reproducible germplasm repositories through sperm cryopreservation is critical for protection and sharing of valuable genetic resources of aquatic organisms that are vital to advancing biomedical research. The Zebrafish International Resource Center (ZIRC) was established to serve as a central repository for genetically modified research lines. In collaboration with the Aquatic Germplasm and Genetic Resources Center (AGGRC) at Louisiana State University Agricultural Center, ZIRC has transitioned successfully from live animal maintenance to high- throughput repositories of cryopreserved sperm. Efficient management of zebrafish genetic lines is extremely important for the research community that uses the zebrafish model to advance biomedical research. Despite this past success, the accelerating number of new research lines is on a path to overwhelm the processing capacity at ZIRC. The reasons for this crisis are rooted in the rapid advancement of genetic editing techniques, and a lack of community-level capabilities to reproducibly cryopreserve sperm in individual laboratories. Less than 1% of zebrafish research laboratories cryopreserve sperm or submit cryopreserved samples to ZIRC. As such, the zebrafish research community has an urgent need for resources to transition from ZIRC-led sperm cryopreservation to a community-driven cryopreservation approach. The goal of the proposed project is to develop a new resource system to provide individual laboratories with decentralized capabilities of reproducible sperm cryopreservation and quality management at the community level. These resources address specific community needs for community-level standards integrated with open hardware to enable reproducible capabilities and training platforms. In contrast to our previous center-focused R-24 projects, this proposed work will employ a user-focused approach, with the principle that any resources developed should be directly accessible, usable, and reproducible by a majority of members of the zebrafish research community. Interdisciplinary innovations will be necessary, including development and integration of 3-D printable open hardware, industrial process management approaches, comprehensive user-ready technology packages, user experience (UX) design methods, and large-scale fabrication and manufacturing. The Specific Aims are to: 1) Establish community-level standards for sample processing and quality management; 2) Develop comprehensive technology packages with open-source hardware to enable community access to decentralized reproducible cryopreservation capabilities, and 3) Develop online resource platforms to engage the user community. These advances will greatly assist ZIRC and the zebrafish community, and will extend beyond the lifetime of the grant, providing a roadmap for germplasm repository development for other biomedical models.