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

非技术段落生物学中的一切都是相互联系的。生命科学家的工作是以最有效、最有效和最经济的方式揭示重要的生物特性。为了做到这一点，他们寻找特别容易处理的模型生物来研究复杂的生物过程，然后应用所学到的知识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，为理解许多基本的生物学过程做出了重大贡献，如受精、胚胎发育和细胞分裂。海胆已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，而且许多特征使实验变得更容易。这笔赠款的目标是创造下一代工具，以加强海胆作为研究模型的效用，使新的研究领域成为可能，并使科学界广泛使用这些工具。从这笔赠款中创建的工具将加强生物学研究的领域，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侮辱和感染，以及组织和器官如何在受损时再生。赠款的一个重要组成部分是超越科学家，接触公众、学生和教师，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。海胆研究人员长期以来一直试图通过遗传方法利用胚胎和成体的实验可控性，但到目前为止，操纵在很大程度上限于对吗啡或药理学的依赖。这笔EDGE赠款的首要目标是建立工具，克服测试棘皮动物基因功能的主要障碍，为跨脊索动物这一宝贵姐妹群体的所有生活史阶段的多样化和综合性研究开辟一个新的发现时代。这一目标将通过优先考虑的重大挑战实现如下：(1)用于基因标记、条件和可逆基因控制的定向DNA插入；(2)用于幼虫培养、变态、少年发育和成人性成熟的快速、标准化的方案，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，以最大限度地利用资源；(4)通过常规的遗传操作，在体外培养胚胎和成体的细胞的简单和高效的方案；(5)常规存档/冷冻保存经过基因操纵的精子、胚胎和细胞，以实现长期可持续性。这些集成了可控和可遗传的遗传操作的新技术，以及在基于细胞的体外系统中测试基因功能和调节的能力，将使充分利用棘皮动物作为研究有机体的重要特性的新的研究途径成为可能。这笔赠款将专注于单一种类的海胆--变异海胆，但这里开发的工具打开的瓶颈将渗透到棘皮动物门中，使其能够在棘皮动物中取得重大进展。将结果快速传播到更大的海胆研究社区是赠款的组成部分。该项目由生物科学局综合组织系统司的开发系统集群(DSC)和既定的刺激竞争研究计划(EPSCoR)共同资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.