Collaborative Research: EDGE FGT: Functional Genomic Tools for Parasitic Nematodes and their Bacterial Symbionts

合作研究：EDGE FGT：寄生线虫及其细菌共生体的功能基因组工具

• 批准号: 2128266
• 负责人: Heidi Goodrich-Blair
• 金额: $ 60万
• 依托单位: University of Tennessee Knoxville
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2128266&HistoricalAwards=false
• 关键词: Collaborative; Research; EDGE; FGT; Functional

All animals, including humans live in association with microbes and parasites that can promote health and cause disease. The mechanisms by which animals communicate with microbes and parasites to block, initiate, maintain, and dissolve such associations are just beginning to be uncovered. Because these mechanisms are often conserved across biology, they can be investigated using model animals such as roundworms, or nematodes, which associate with microbes and are themselves parasites. This project will develop new experimental tools in roundworms that have developed partnerships, or symbioses, with specific bacteria. Together these roundworms and their bacterial partners infect and kill insects, using them as a food source. Developing new tools to study this elegant animal-bacterium system will help expand our understanding how animals and bacteria form partnerships, and how they work together to parasitize other animals. The tools and knowledge gained in this project will be rapidly shared with the community of researchers involved in drug discovery, agricultural control of crop pests, and in the study of parasitism, infectious disease, beneficial microbiome function, and fundamental cell, molecular, developmental, and evolutionary biology. As part of this project, undergraduates will be involved in a discovery-based microbiology lab where they will practice isolating and identifying new insect-killing roundworms from the environment. Young people (K-12) and educators will be engaged through collaboration with the Science Journal for Kids, where a basic curriculum and summary of key research findings will be developed for classroom use. Entomopathogenic nematodes in the genera Heterorhabditis and Steinernema are mutualistically associated with bacteria in the genera Photorhabdus and Xenorhabdus, respectively. The nematode-bacterium symbiotic pair obligately parasitizes insects as a nutrient source for reproduction and has utility as a biological control agent for agricultural insect pests and as a source of novel compounds. The entomopathogenic pair and their insect hosts are models to understand fundamental biological principles, including the evolution and molecular and cellular basis of mutualistic and antagonistic organismal interactions. Numerous features make this system an excellent experimental model including ease and cost of husbandry, fast generation time, and optical transparency. Although genetic techniques have been developed for representative bacterial symbiotic partners and insect hosts, to date there have been no broadly adopted, reliable genetic modification tools in either Steinernema or Heterorhabditis nematodes. This inability to interrogate nematode gene function has hobbled full use of this system to yield much-needed insights into parasitism, animal microbiome interactions, and other areas. Here we propose to capitalize on a recently isolated Steinernema nematode that has promising characteristics for development of genetic tools, including hermaphroditic reproduction, amenability to long-term freezing, healthy development on agar bacterial lawns, resilience to microinjection, and significant pathogenicity to lab insects and agricultural pests. By fully sequencing the genomes and developing genetic techniques and tools for both nematode and symbiont, including CRISPR-Cas9 genome editing in the nematode, and creating an arrayed mutant library for the bacterium, our team will help realize the full potential of this elegant animal-microbe model system.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.

包括人类在内的所有动物都与微生物和寄生虫结合，可以促进健康并引起疾病。动物与微生物和寄生虫沟通以阻止，启动，维持和溶解此类关联的机制刚刚开始被发现。由于这些机制通常是在生物学中保守的，因此可以使用模型动物（例如round虫或线虫）进行研究，它们与微生物相关，本身就是寄生虫。该项目将开发与特定细菌建立伙伴关系或共生伙伴关系的新型实验工具。这些round虫及其细菌伴侣将它们作为食物来源感染并杀死昆虫。开发研究这种优雅的动物 - 细菌系统的新工具将有助于扩大我们的理解方式，以及它们如何形成伙伴关系，以及它们如何共同努力以使其他动物寄生。该项目中获得的工具和知识将与参与药物发现的研究人员，农作物害虫的农业控制以及寄生虫，传染病，有益的微生物组功能以及基本细胞，分子，发育和进化生物学的研究迅速分享。作为该项目的一部分，本科生将参与一个基于发现的微生物学实验室，他们将练习与环境中隔离和识别新的杀死昆虫的round虫。年轻人（K-12）和教育工作者将通过与儿童科学杂志的合作进行，其中将开发基本的课程和关键研究结果的摘要，以供课堂使用。异源性疾病中的昆虫致病线虫和施泰氏菌分别与Photorhabdus属和Xenorhabdus中的细菌相互关联。线虫 - 细菌共生对有必要将昆虫寄生为繁殖的营养来源，并具有效用作为农用虫害的生物控制剂，并且是新型化合物的来源。昆虫病态对及其昆虫宿主是了解基本生物学原理的模型，包括相互和拮抗生物体相互作用的进化以及分子和细胞基础。许多功能使该系统成为出色的实验模型，包括畜牧业的易于和成本，快速生成时间和光学透明度。尽管已经为代表性的细菌共生伴侣和昆虫宿主开发了遗传技术，但迄今为止，在Steinernema或Heterorhabditis nematodes中尚无广泛采用的可靠的遗传修饰工具。这种无法审问线虫基因功能的无法充分利用该系统，以产生对寄生虫，动物微生物组相互作用和其他领域的急需的见解。在这里，我们建议利用最近孤立的Steinernema线虫，该线虫具有有望开发遗传工具的特征，包括雌雄同体的繁殖，长期冻结的适应性，对琼脂细菌草坪的健康发展，对微分射的韧性以及对实验室昆虫和农业虫害的强大的致病性。通过对线虫和共生体的开发基因组并开发遗传技术和工具，包括CRISPR-CAS9基因组在线虫中编辑，并为细菌创建一个阵列的突变图库，我们的团队将帮助实现这一优雅的动物模型的全部潜力。更广泛的影响审查标准。