CAREER: Characterizing germ granule diversity and its influence on germline development in Drosophila

动物繁殖需要高度特化的细胞，这些细胞可以将遗传信息传递给下一代。这些细胞统称为生殖细胞，含有称为生殖颗粒的细胞结构，其中包含负责确保生殖细胞功能的不同类型的生物分子。该项目的总体研究目标是了解胚芽颗粒的含量如何改变以影响动物的发育和繁殖。拟议的实验将在包括教育和研究整合计划，建立新泽西的研究联盟，为包容性的干教育进行。这个新的三阶段计划旨在通过当地高中，基恩大学和普林斯顿大学之间的多机构合作，为代表性不足的群体提供研究机会。该提案提供了显微镜培训和一个利用生物图像促进整个社区对科学的热情的推广计划。该项目旨在揭示影响动物繁殖的机制，支持多样化和有竞争力的科学劳动力的发展，通过实践研究培训和社区参与改善教育，并作为高中和研究型大学之间正式合作的模式。不同果蝇物种在生殖颗粒内存在的某些转录物的丰度方面表现出显着的多样性。该项目旨在确定控制生殖颗粒多样性的机制，并探索这些变化如何影响生殖细胞的发育和繁殖。PI假设胚芽颗粒mRNA含量的多样性受三种机制的协同调节：mRNA表达水平的差异，胚芽颗粒mRNA携带能力的变化，以及mRNA掺入颗粒的功效的变化。 目的1进一步表征胚芽颗粒的多样性，通过定量10个基因的mRNA在单个颗粒中的丰度，并使用单分子原位杂交和定量成像在10个果蝇物种中进行比较。目的2结合生物学数据和计算模型来测量每个机制对胚芽颗粒mRNA含量的影响。转录本在胚粒中的作用受转录本3′ UTR顺式作用元件的调控。使用基因组编辑，目的3研究3′ UTR内的进化差异是否可以改变生殖颗粒mRNA含量和种系发育。不同种类的颗粒具有相似的成分，该项目的发现可能对其他类型的生物分子凝聚物有更广泛的影响。该奖项是共同的，该奖项由分子和细胞生物科学部的遗传机制计划和综合有机体系统部的发育系统计划资助。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的学术价值和更广泛的影响审查标准。

Animal reproduction requires highly specialized cells that can pass on genetic information to the following generation. These cells, collectively known as the germline, contain cellular structures called germ granules, which house different types of biomolecules that are responsible for ensuring germline function. The overall research goal for this project is to understand how the contents of germ granules can be changed to influence animal development and reproduction. The experiments proposed will be performed within the included Education and Research Integration Plan which establishes New Jersey’s Research Alliance for Inclusive STEM Education. This new, three-phase program is designed to elevate research opportunities for underrepresented groups via multi-institution collaboration between local high schools, Kean University, and Princeton University. The proposal offers microscopy training and an outreach plan that uses biological images to promote enthusiasm about science throughout the community. Together, this project aims to shed light on mechanisms that can influence animal reproduction, supports the development of a diverse and competitive scientific workforce, improves education with hands-on research training and community engagement, and serves as a model for formalizing partnerships between high schools and research universities.Different Drosophila species display remarkable diversity in the abundance of certain transcripts that reside within germ granules. This project seeks to identify the mechanisms that control germ granule diversity and explore how such variations influence germline development and reproduction. The PI hypothesizes that diversity in germ granule mRNA content is cooperatively regulated by three mechanisms: differences in mRNA expression levels, variations in germ granules’ mRNA carrying capacity, and changes to the efficacy with which mRNAs become incorporated into granules. Aim 1 further characterizes germ granule diversity by quantifying the abundance in individual granules of mRNA from ten genes and comparing them across ten Drosophila species using single molecule in situ hybridization and quantitative imaging. Aim 2 combines biological data with computational modeling to measure the impact of each mechanism on germ granule mRNA content. The efficacy with which transcripts populate germ granules can be regulated by cis-acting elements in the 3′ UTR of each transcript. Using genome editing, Aim 3 investigates whether evolutionary differences within 3′ UTRs can alter germ granule mRNA content and germline development. Different classes of granules share similar components, and discoveries from this project may have broader implications for other types of biomolecular condensates.This award is co-funded by the Genetic Mechanisms Program in the Molecular and Cellular Biosciences Division and the Developmental Systems Program in the Integrative Organismal Systems Division.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.