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.

项目摘要/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 每年通过多种方法，包括诱变、基因转移、基因编辑、基因敲除、 在 NIH 资助的五个水产资源中心中，单倍体的保存。 种质（精子）可用于三个群体：斑马鱼、剑鱼和爪蟾。 没有实用的方法可用于保存任何二倍体种质（例如胚胎或幼虫） 这个问题对于海兔（Aplysia californica）来说尤其突出，因为它是一种海兔。 生殖特征（非自体受精雌雄同体）需要低温保存早期生命阶段 而不是精子，冷冻二倍体种质的可用性将大大加速。 通过消除筛选和生产多个研究线的需求 因此，社区需要保存和使用遗传。 通过新技术，所有水生生物医学模型的资源将大大增加 胚胎和幼虫的冷冻保存，这项技术对于海兔来说是必不可少的，因此，我们的目标是。 开发新型电冷冻生物学技术来监测和改善玻璃化和冷冻恢复 水生生物医学模型二倍体基因组的方案具体目标是：1）开发新颖的。 与海兔相关的低温过程超快速监测（毫秒）技术 胚胎玻璃化冷冻；2）利用新颖的监测技术获得对传统胚胎玻璃化冷冻的新见解。 海兔胚胎玻璃化冷冻的方法，以及 3) 扩展监测技术以允许交互 尽管这项工作通过低温过程获得了海兔胚胎玻璃化冷冻的新方法。 专注于海兔作为补充剂，可用于青蛙、火蜥蜴的胚胎和精子， 和鱼类，并将为未来扩大公用事业和鱼类的赠款提案提供所需的初步数据 通过添加该补充材料，对于推进家长奖励将非常有价值。 强大的新方法（目标 1）将极大地加速我们当前的传统研究（目标 2），它将为我们提供新的工具来建立新的方法（目标 3）。 其他 R24 资金）使我们能够组建一个独特的跨学科团队，拥有超过 15 年的经验 这项工作各个方面的专业知识，涵盖生殖生物学、低温生物学、工程和设备 这项工作可以在当前项目年度（进入第 3 年）内完成，但不能。 与家长奖励或之前的补充资金资助的工作重叠。