EDGE CT: Tools to advance functional genomic studies in sea urchins

生物学中的一切都是相互联系的。以最具影响力、最有效和最经济的方式揭示重要的生物特性是生命科学家的工作。为了做到这一点，他们寻找特别容易处理的模式生物，用于研究复杂的生物过程，然后应用所学到的知识来更好地了解其他生物。一个多世纪以来，海胆提供了一个有价值的研究模型，为理解受精、胚胎发育和细胞分裂等许多基本生物学过程做出了重大贡献。海胆被证明是一个有价值的模型，因为它们与脊椎动物有着密切的遗传关系，而且它们的许多特征使实验更容易。这项拨款的目标是创建下一代工具，以增强海胆作为研究模型的效用，从而使新的研究领域成为可能，并使这些工具广泛地为科学界所用。由这笔拨款创建的工具将加强生物学研究领域，包括更好地了解卵子和精子在受精时如何相互作用，了解胚胎发育的规则，神经细胞是如何形成的，性别是如何决定的，动物如何保护自己免受环境的伤害和感染，以及组织和器官在受损时如何再生。该基金的一个重要组成部分是让公众、学生和教师都能接触到科学家，并使海胆成为21世纪极具吸引力和影响力的研究和教育工具。长期以来，海胆研究人员一直试图利用基因方法来利用胚胎和成体海胆的实验可操控性，但迄今为止，操作在很大程度上仅限于依赖于形态学或药理学。这项EDGE资助的总体目标是建立工具，克服测试棘皮动物基因功能的主要障碍，为这一有价值的脊索动物姐妹群体的所有生活史阶段的多样化和综合研究开辟一个新的发现时代。这一目标将通过以下优先级大挑战来实现：(1)靶向DNA插入，用于基因标记，条件和可逆的基因控制；(2)快速、标准化的幼虫培养、变态、幼鱼发育和成虫性成熟方案，包括每个阶段的营养和环境优化；(3)快速、简单的动物性别鉴定方案，使资源最大化；(4)简单有效的胚胎和成体细胞体外常规遗传操作培养方案；(5)常规存档/冷冻保存基因操纵的精子、胚胎和细胞，以实现长期可持续性。这些综合的新技术与可控和可遗传的基因操作以及在体外细胞系统中测试基因功能和调控的能力将为充分利用棘皮动物作为研究生物的重要特性提供新的研究途径。这笔拨款将集中于一种海胆，Lytechinus variegatus，但这里开发的工具打开的瓶颈将渗透到整个门，使棘皮动物取得重大进展。将研究结果迅速传播给更大的海胆研究人员群体是该基金的一个组成部分。该项目由生物科学理事会综合有机体系统部的发展系统集群（DSC）和刺激竞争研究的既定计划（EPSCoR）共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Non-Technical Paragraph Everything in biology is connected. It is the job of life scientists to reveal important biological properties in the most impactful, efficient, and economical way. To do so they look for model organisms that are particularly tractable for studying complex biological processes and then apply what is learned to better understand other organisms. For more than a century, sea urchins have provided a valuable research model that has contributed significantly to understanding of many fundamental biological processes such as fertilization, embryonic development and cell division. Sea urchins have proven to be a valuable model due to their close genetic relationship to vertebrate animals and many features that make experimentation easier. The goal of this grant is to create the next generation of tools to enhance the utility of sea urchins as research models that will enable new areas of research and to make these tools widely available to the scientific community. Areas of biological research to be enhanced by the tools created from this grant include a better understanding of how eggs and sperm interact at fertilization, understanding the rules of embryo development, how nerve cells are made, how sex is determined, how animals protect themselves from environmental insults and from infection, and how tissues and organs can regenerate when they are damaged. An essential component of the grant is to reach beyond scientists to the public, students and teachers and to make the sea urchin a highly attractive and impactful research and education tool of the twenty-first century. Technical Paragraph Sea urchin researchers have long sought to leverage the experimental tractability of the embryo and adult with genetic approaches but, to date, manipulations have been limited largely to dependence on morpholinos or pharmacology. The overarching goal of this EDGE grant is to build tools that overcome major obstacles to testing gene functionality in echinoderms, opening up a new era of discovery for diverse and integrated studies across all life history stages of this valuable sister group to chordates. This goal will be realized by prioritized Grand Challenges as follows: (1) Targeted DNA insertion for gene tagging, conditional, and reversible gene control; (2) Rapid, standardized protocols for larval culturing, metamorphosis, juvenile development, and adult sexual maturation including nutritional and environmental optimization for each stage; (3) Rapid, simple sexing protocols of animals to maximize resources; (4) Simple and efficient protocols for culturing cells from the embryo and adult in vitro with routine genetic manipulations; (5) Routine archiving/cryopreserving genetically manipulated sperm, embryos and cells for long term sustainability. These integrated new technologies with controlled and heritable genetic manipulations and the ability to test gene function and regulation in in vitro cell-based systems will enable new avenues of investigation that fully exploit the important properties of echinoderms as a research organism. This grant will focus on a single species of sea urchin, Lytechinus variegatus, but the bottlenecks opened by tools developed herein will permeate the phylum enabling significant advances across echinoderms. Rapid dissemination of results to the larger community of sea urchin researchers is an integral part of the grant.This project is jointly funded by the Developmental Systems Cluster (DSC) in the Division of Integrative Organismal Systems of the Biological Sciences Directorate and the Established Program to Stimulate Competitive Research (EPSCoR).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.