CAREER: Investigating the Role of an RNA Interference Pathway in Safeguarding the Tetrahymena Thermophila Somatic Genome

该奖项的全部或部分资金来自《2021年美国救援计划法案》(公法117-2)。保护我们的基因组免受突变和DNA损伤是维持生命和预防疾病的关键。虽然细胞确保基因组完整性的方法有多种，但最近的研究揭示了由RNA分子和RNA相关蛋白质组成的基因组保护的新参与者。然而，关于这些生物分子是如何发挥作用的，仍有许多需要了解。因此，该项目的目标是通过研究一种单细胞生物体中基于RNA的过程来加深我们对RNA及其相关蛋白质如何确保基因组完整性的理解。长期以来，该过程一直是生物学基础发现的基础：嗜热四膜虫。该项目提供的科学见解预计将对包括人类在内的各种生物如何使用基于RNA的过程来保护它们的基因组以及这些过程的中断可能如何导致疾病产生重要影响。此外，该项目将为西北华盛顿大学、传统大学、太平洋大学和卡罗尔学院四所主要本科机构的本科生提供机会，扩大获得真实研究经验和STEM专业发展的机会。由于这些机构服务于一些在STEM中被认为是代表不足的群体的学生，该项目的教育方面预计将促进不同学生在科学领域的保留和成功，从而加强我们未来的科学工作。从细胞分裂和分化、抵御病毒和转座子，到生物发育，各种生物过程都依赖于RNA干扰(RNAi)途径。虽然RNAi被认为是一种抑制基因表达并确保着丝粒周围适当染色质结构的机制，但最近在各种真核生物中的研究发现，RNAi因子在防止或修复DNA损伤方面发挥了新的作用。这些新角色的细节以及它们如何在进化上得到保守，仍有待充分阐明。这个项目将研究RNAi在纤毛模式真核生物嗜热四膜虫中维持基因组完整性的新发现的作用的机制和基因组定位。这项工作将利用已建立的反向遗传、荧光显微镜和生化方法，开发或调整用于四膜虫。这些研究有可能揭示RNAi管理的基因组完整性的特定物种和广泛保守的机制，促进我们对RNA在基因组保护中新出现的角色的理解。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This award is funded in whole or in part under the American Rescue Plan Act of 2021 (Public Law 117-2).Protection of our genomes from mutation and DNA damage is critical for sustaining life and preventing disease. While there are a variety of ways by which cells ensure genome integrity, recent studies have revealed new players in genome protection that consist of RNA molecules and RNA-associated proteins. Yet much remains to be understood about how these biomolecules function. Therefore, the goal of this project is to deepen our understanding of how RNA and RNA-associated proteins ensure the integrity of genomes through investigation of an RNA-based process in a single-celled organism that has long served as the basis of foundational discoveries in biology: Tetrahymena thermophila. The scientific insights afforded by this project are expected to have important implications for how diverse organisms, including humans, use RNA-based processes to protect their genomes and how disruptions to these processes may underlie disease. In addition, this project will provide opportunities to expand access to authentic research experiences and STEM professional development for undergraduate students at four primarily undergraduate institutions in the Northwest – Western Washington University, Heritage University, Pacific University, and Carroll College. As these institutions serve a number of students who identify as members of underrepresented groups in STEM, the education aspects of this project are anticipated to promote the retention and success of diverse students in science, thereby strengthening our future scientific workforce.Biological processes as diverse as cell division and differentiation, defense against viruses and transposons, and organismal development depend on RNA interference (RNAi) pathways. While RNAi is best understood as a mechanism that represses gene expression and ensures proper chromatin structure around centromeres, recent studies in diverse eukaryotes have uncovered new roles for RNAi factors in preventing or repairing DNA damage. The details of these new roles and how evolutionarily conserved they are remain to be fully elucidated. This project will investigate the mechanisms and genomic locations for a newly uncovered role for RNAi in maintaining genome integrity in the ciliated model eukaryote Tetrahymena thermophila. This work will leverage established reverse genetic, fluorescence microscopy, and biochemical approaches developed or adapted for use in Tetrahymena. These studies have the potential to reveal both species-specific and broadly conserved mechanisms for RNAi-governed genome integrity, advancing our understanding of emerging roles for RNA in genome protection.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.