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

EDGE CT：推进海胆功能基因组研究的工具

• 批准号: 1923445
• 负责人: Gary Wessel
• 金额: $ 128.49万
• 依托单位: Brown University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1923445&HistoricalAwards=false
• 关键词: EDGE; CT; Tools; advance; functional

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.

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学特性。为了做到这一点，他们寻找特别易于研究复杂生物过程的模式生物，然后将所学到的知识应用于更好地了解其他生物。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过程，如受精，胚胎发育和细胞分裂做出了重大贡献。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，并且具有许多使实验更容易的特征。 这笔赠款的目标是创建下一代工具，以提高海胆作为研究模型的实用性，这将使新的研究领域成为可能，并使这些工具广泛提供给科学界。生物学研究领域将通过这项赠款创建的工具得到加强，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，神经细胞如何形成，性别如何决定，动物如何保护自己免受环境侵害和感染，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，不仅要让科学家接触到公众、学生和教师，而且要使海胆成为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胚胎和成体的实验易处理性与遗传方法，但迄今为止，操作主要限于依赖吗啉代或药理学。该EDGE赠款的总体目标是建立克服棘皮动物基因功能测试主要障碍的工具，为这个有价值的姐妹群体脊索动物的所有生活史阶段的多样化和综合研究开辟了一个新的发现时代。这一目标将通过以下优先大挑战来实现：（1）用于基因标签、条件和可逆基因控制的靶向DNA插入;（2）用于幼虫培养、变态、幼年发育和成体性成熟的快速、标准化方案，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）通过常规遗传操作体外培养胚胎和成人细胞的简单有效方案;（5）常规存档/冷冻保存遗传操作的精子、胚胎和细胞，以实现长期可持续性。这些新技术与可控和可遗传的遗传操作以及在体外基于细胞的系统中测试基因功能和调节的能力相结合，将使新的研究途径能够充分利用棘皮动物作为研究生物体的重要特性。这项资助将集中在海胆的一个单一物种，Lytechinus variegatus，但这里开发的工具打开的瓶颈将渗透到整个棘皮动物门，使其取得重大进展。 该项目由生物科学理事会综合有机系统司的发展系统组（DSC）和刺激竞争研究的既定计划（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。

项目成果

Optimizing CRISPR/Cas9-based gene manipulation in echinoderms.

优化棘皮动物中基于 CRISPR/Cas9 的基因操作。

• DOI: 10.1016/j.ydbio.2022.07.008
• 发表时间: 2022
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Oulhen N;Pieplow C;Perillo M;Gregory P;Wessel, GM
• 通讯作者: Wessel, GM