Collaborative Research: EDGE FGT: Genome-wide knock-out mutant libraries for the microcrustacean Daphnia

合作研究：EDGE FGT：微甲壳动物水蚤的全基因组敲除突变体文库

• 批准号: 2324639
• 负责人: Sen Xu
• 金额: $ 110万
• 依托单位: University of Missouri-Columbia
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2324639&HistoricalAwards=false
• 关键词: Collaborative; Research; EDGE; FGT; Genome

Mutating the DNA sequences of genes of interest is a valuable approach for understanding their biological functions in diverse organisms. In contrast to established model systems, large collections of gene knock-out mutants are not widely available in emerging model organisms. This limitation greatly diminishes the power of emerging model systems for making novel scientific discoveries. To address the issue, this project will develop a large collection of gene knock-out mutants using CRISPR gene editing in three species of the freshwater microcrustacean Daphnia (D. magna, D. pulex, and D. pulicaria), which is an emerging model system in ecological genomics, evolutionary genomics, and environmental genomics. The Daphnia knock-out mutants will provide unprecedented opportunities for the scientific community to characterize the biological functions of the genes. The project will also disseminate technical expertise for CRISPR gene editing in Daphnia through free workshops, targeting biologists at different career stages including undergraduates, graduates, postdoctoral researchers, and faculty. Furthermore, the project will develop an innovative genomics curriculum to enable hands-on research in the classroom and enhance the education experience of diverse undergraduate students. The plan is to knock out genes in Daphnia, including many candidate genes that may play critical roles in acclimation, adaptation, and evolution of these species. Individual knock-out mutant lines will be generated, maintained in the laboratory, and freely shared with the research community. Each mutant line will be whole-genome sequenced to characterize its genomic background. In collaboration with the global Daphnia research community, this project will evaluate the functional changes in knock-out mutants at both genomic and organism levels to validate the role of the genes in various environmental settings and to infer gene interaction and gene regulation networks. Lastly, a free, public-facing database will be developed to host the genomic and phenotypic data of the mutant lines, to promote and manage the distribution of Daphnia mutants, and to distribute experimental protocols and reagents.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.

突变感兴趣的基因的DNA序列是理解其在多种生物中的生物学功能的宝贵方法。与已建立的模型系统相反，在新兴模型生物体中，大量基因敲除突变体并未广泛使用。这种局限性大大削弱了新兴模型系统在做出新的科学发现方面的力量。为了解决这个问题，该项目将使用CRISPR基因编辑来开发大量的基因基因敲除突变体，这是三种淡水Microcrustacean daphnia（D。Magna，D。Pulex和D. plicaria）的三种物种，这是生态基因组学，进化基因组学和环境基因组学的新兴模型系统。水庄的敲除突变体将为科学界提供前所未有的机会来表征基因的生物学功能。该项目还将通过免费的研讨会来传播水壶中CRISPR基因编辑的技术专业知识，针对不同职业阶段的生物学家，包括本科生，毕业生，博士后研究人员和教职员工。此外，该项目将开发创新的基因组学课程，以便在课堂上进行动手研究，并增强多元化本科生的教育经验。 该计划是淘汰水坝中的基因，其中包括许多可能在这些物种的适应，适应和进化中起关键作用的候选基因。将在实验室中产生，维护单独的淘汰突变线，并与研究界自由共享。每个突变系列将是全基因组测序的，以表征其基因组背景。该项目与全球水白原研究界合作，评估基因组和生物体水平上的基因敲除突变体的功能变化，以验证基因在各种环境环境中的作用，并推断基因相互作用和基因调节网络。最后，将开发一个免费的，面向公共的数据库，以托管突变线的基因组和表型数据，以促进和管理流浪者突变体的分布，分发实验方案和试剂。这奖反映了NSF的法规任务，并被认为是通过基金会的知识优点和宽阔的范围来评估的，并且值得通过评估来进行评估。