Safeguarding Genetic Resources of Aquatic Biomedical Models - Research Supplemental

保护水生生物医学模型的遗传资源 - 研究补充

• 批准号: 10609341
• 负责人: Maria Teresa Gutierrez-Wing
• 金额: $ 27.42万
• 依托单位: LOUISIANA STATE UNIV AGRICULTURAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10609341
• 关键词: Aplysia; Aplysia species; Applications Grants; Award; Backcrossings; Communities; Cryopreservation; Crystallization; Data; Devices; Diploidy; Embryo; Engineering; Fishes; Freezing; Funding; Future; Gene Transfer; Generations; Genes; Genome; Goals; Haploidy; Hermaphroditism; Ice; Intention; Larva; Life; Methods; Modeling; Monitor; Monoclonal Antibody R24; Mutagenesis; Parents; Process; Production; Protocols documentation; Rana; Recovery; Reproductive Biology; Research; Salamander; Technology; United States National Institutes of Health; Water; Work; Xenopus; Xiphophorus; Zebrafish; cell injury; cryobiology; cryogenics; electric field; embryo cryopreservation; genetic resource; improved; insight; knockout gene; millisecond; new technology; novel; novel strategies; preservation; reproductive; screening; sperm cell; tool; trait

Project Summary/Abstract: Within aquatic biomedical models, thousands of valuable research lines are created and characterized each year by numerous methods including mutagenesis, gene transfer, gene editing, gene knockout, hybridization, and backcrossing. Among the five NIH‐funded aquatic stock centers, preservation of haploid germplasm (sperm) is possible for three groups: zebrafish, Xiphophorus and Xenopus. Currently there is no practical method available for preservation of diploid germplasm (e.g., embryos or larvae) for any aquatic group. This problem is especially prominent for the sea hare, Aplysia californica, which because of reproductive traits (non‐self‐fertilizing hermaphroditism) requires cryopreservation of early life stages rather than sperm. The availability of cryopreserved diploid germplasm would greatly accelerate the availability of numerous research lines by removing needs for screening and production of multiple generations to produce homozygosity. Thus, the community needs for preservation and use of genetic resources would be greatly advanced for all aquatic biomedical models by novel technologies for cryopreservation of embryos and larvae, and this technology is essential for Aplysia. Therefore, our goal is to develop novel electro‐cryobiology technologies to monitor and improve vitrification and cryo‐recovery protocols for diploid genomes of aquatic biomedical models. The Specific Aims are to: 1) develop novel technologies for ultra‐rapid monitoring (millisecond) of cryogenic processes associated with Aplysia embryo vitrification; 2) use the novel monitoring technology to gain new insights into traditional approaches for Aplysia embryo vitrification, and 3) expand the monitoring technology to allow interaction with cryogenic processes to gain new approaches for Aplysia embryo vitrification. This work, although focused on Aplysia for the supplement, would be useful for embryos and sperm of frogs, salamanders, and fishes, and would provide preliminary data needed for future grant proposals to expand utility and application. This supplement would be extremely valuable in advancing the parent award by adding powerful new approaches (Aim 1) that would greatly accelerate our current conventional research (Aim 2), and it would provide us with novel tools to establish new approaches (Aim 3). The parent award (and other R24 funding) has allowed us to assemble a unique interdisciplinary group with more than 15 years of expertise in all aspects of this work spanning reproductive biology, cryobiology, engineering, and device fabrication. This work can be completed within the current project year (entering Year 3), and it does not overlap with work funded in the parent award or through previous supplemental funding.

项目概要/摘要： 在水生生物医学模型中，创建并表征了数千条有价值的研究线 每年通过许多方法，包括诱变，基因转移，基因编辑，基因敲除， 杂交和回交。在NIH资助的五个水产资源中心中， 种质（精子）可能用于三个群体：斑马鱼、剑尾鱼和非洲爪蟾。目前还 没有可用于保存二倍体种质的实用方法（例如，胚胎或幼虫） 水生类群。这一问题对于海兔，加利福尼亚海兔，尤其突出，因为 生殖特征（非自体雌雄同体）需要冷冻保存早期生命阶段 而不是精子。超低温保存的二倍体种质的利用将大大促进 通过消除筛选和生产多种产品的需求，提供众多研究线 产生纯合性。因此，社区需要保存和利用遗传资源， 资源将大大先进的所有水生生物医学模型的新技术， 冷冻保存胚胎和幼虫，这项技术是必不可少的。因此，我们的目标是 开发新的电低温生物学技术，以监测和改善玻璃化和冷冻复苏 水生生物医学模型的二倍体基因组方案。具体目标是：1）开发新的 超快速监测技术（毫秒）与低温相关的低温过程 胚胎玻璃化; 2）使用新的监测技术，以获得新的见解， 方法为Astrassia胚胎玻璃化，和3）扩大监测技术，以允许相互作用 利用低温冷冻技术，为亚洲红豆杉胚胎玻璃化冷冻提供新的途径。这项工作，虽然 专注于Aaplasia的补充，将是有用的胚胎和精子的青蛙，蝾螈， 和鱼类，并将提供未来赠款提案所需的初步数据，以扩大效用， 应用程序.这一补充将是非常宝贵的，在推进父母的裁决，增加 强大的新方法（目标1），将大大加快我们目前的传统研究（目标 2），它将为我们提供新的工具来建立新的方法（目标3）。家长奖（及 其他R24资金）使我们能够组建一个独特的跨学科小组， 在这项工作的各个方面，包括生殖生物学、低温生物学、工程学和设备学， 捏造这项工作可以在本项目年度（进入第3年）内完成， 与母基金资助的工作或通过以前的补充资金重叠。