Resources for Drosophila embryo cryopreservation at lab and stock center scale

实验室和库存中心规模的果蝇胚胎冷冻保存资源

• 批准号: 10569277
• 负责人: JOHN C BISCHOF
• 金额: $ 74.57万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10569277
• 关键词: Acceleration; Adoption; Adult; Animal Model; Benchmarking; Biomedical Research; Collaborations; Collection; Communities; Cost-Benefit Analysis; Costs and Benefits; Cryopreservation; Disasters; Drosophila genus; Drosophila melanogaster; Educational workshop; Embryo; Event; Feedback; Freezing; Future; Genetic; Genetic Diseases; Genetic Drift; Goals; Human Resources; Indiana; Individual; Larva; Maintenance; Malignant Neoplasms; Methods; Outcome; Outcomes Research; Pathway interactions; Population; Prevention; Procedures; Protocols documentation; Reagent; Reproducibility; Research; Research Personnel; Resource Sharing; Resources; Rewarming; Risk; Services; Shipping; Source; Standardization; Temperature; Testing; Time; Training; Work; cost; cost effective; cost estimate; cryobiology; cryogenics; embryo cryopreservation; experience; flexibility; fly; genetic resource; hatching; innovation; instrument; next generation sequencing; preservation; prevent; research and development; success; tool

ABSTRACT Drosophila melanogaster is one of the leading animal models for biomedical research, with researchers generating new lines of Drosophila every year for studies in numerous cancers, genetic disorders, and other maladies. Currently there are greater than 160,000 Drosophila stocks held at different stock centers around the world. As reliable and cost-effective approaches for long-term preservation of Drosophila stocks are lacking, individual labs and stock centers must maintain their lines as living populations, which is resource-intensive and puts the stocks at risk of loss. In 2021, our group developed an easily implemented and robust cryopreservation protocol for Drosophila melanogaster embryos which can be applied in any lab without the need for specialized instruments. This protocol is broadly applicable, and it has been successfully used to preserve 25 distinct strains from different sources. For most strains, >50% of the embryos hatch and >25% of the resulting larvae develop into fully functioning adults after cryopreservation and rewarming (normalized survival to control embryos), providing sufficient numbers of adults to revive the strain. To accelerate the cryopreservation and archival of critical Drosophila stocks, we will develop, standardize, and disseminate resources to individual labs and stock centers. Significant efforts will be devoted to developing reliable methods for cryogenic storage and shipping of Drosophila embryos as well as to study if any significant mutagenic changes occur due to cryopreservation. We will be working with multiple groups within the cryobiology and fly community to gather feedback on and refine our approach. With the collaboration of the Bloomington Drosophila Stock Center (BDSC), we will identify and prioritize the stocks that need cryopreservation to prevent their total loss in the event of a disaster or genetic drift. We will also demonstrate the efficacy of our cryopreservation protocol on a large scale with a selected group of stocks identified by BDSC. Finally, we will also estimate the costs for cryopreservation in comparison to the traditional method of Drosophila maintenance to inform future efforts to protect these precious genetic resources. This is a multi-PI project led by investigators with significant experience in Drosophila genetics and cryobiology.

抽象的 果蝇Melanogaster是生物医学研究的主要动物模型之一，研究人员 每年生成新的果蝇线，以研究众多癌症，遗传疾病和其他 疾病。目前，在周围不同股票中心持有的果蝇股票超过160,000股 世界。由于缺乏可靠且具有成本效益的长期保护果蝇股票的方法，因此 各个实验室和股票中心必须将其界限作为生活人群，这是资源密集的， 使股票处于损失的危险之中。在2021年，我们的小组开发了一种易于实施且强大的冷冻保存 果蝇果蝇的协议，可以在任何实验室中应用，而无需专业 乐器。该协议广泛适用，已成功用于保留25种不同的应变 来自不同的来源。对于大多数菌株，> 50％的胚胎孵化和> 25％的幼虫发展 在冷冻保存和重新加热后进入功能齐全的成年人（对控制胚胎的归一化生存）， 提供足够数量的成年人来恢复菌株。加速冷冻保存和档案 关键的果蝇股票，我们将开发，标准化和传播资源 中心。重大努力将致力于开发可靠的低温存储和运输方法 果蝇胚胎以及研究是否由于冷冻保存而发生任何明显的诱变变化。我们 将与冷冻生物学和飞行社区中的多个小组合作，以收集反馈和完善 我们的方法。通过布卢明顿果蝇股票中心（BDSC）的合作，我们将确定和 优先考虑需要冷冻保存的股票，以防止发生灾难或遗传的股票 漂移。我们还将大规模证明我们的冷冻保存协议的功效 BDSC确定的一组股票。最后，我们还将估计冷冻保存的成本 果蝇维护的传统方法，以告知未来的努力来保护这些宝贵的遗传 资源。这是一个由研究人员领导的多PIP项目，在果蝇遗传学和 冷冻生物学。