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，包含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如何改变胚芽颗粒的含量来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内进行，该计划建立了新泽西州的全纳STEM教育研究联盟。这个新的三阶段项目旨在通过当地高中、肯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增加研究机会。该提案提供了显微镜培训和一项推广计划，该计划使用生物图像来促进整个社区对科学的热情。总之，该项目旨在揭示影响动物繁殖的机制，支持发展多样化和有竞争力的科学劳动力，通过实践研究培训和社区参与改善教育，并成为高中和研究型大学之间正式建立伙伴关系的典范。不同种类的果蝇在胚芽颗粒内某些转录本的丰度上表现出显著的多样性。该项目旨在确定控制胚粒多样性的机制，并探索这种变异如何影响种系发育和生殖。PI假设，胚粒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、胚粒mRNA携带能力的变化以及mRNA被纳入颗粒的效力的变化。目的1通过量化来自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技术对10个果蝇物种的mRNA进行比较，进一步表征了细菌颗粒的多样性。目的2将生物学数据与计算模型相结合，测量每种机制对胚粒mRNA含量的影响。转录物填充胚芽颗粒的效果可由每个转录物3 ' UTR中的顺式作用元件调节。利用基因组编辑，Aim 3研究了3 ' utr内的进化差异是否可以改变胚粒mRNA含量和种系发育。不同类型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析物具有更广泛的意义。该奖项由分子和细胞生物科学部的遗传机制项目和综合有机系统部的发育系统项目共同资助。该奖项反映了美国国家科学基金会的法定使命，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。

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.