Mechanisms of Small RNA-Mediated Silencing in Chlamydomonas

衣藻中小 RNA 介导的沉默机制

• 批准号: 2131783
• 负责人: Heriberto Cerutti
• 金额: $ 79.48万
• 依托单位: University of Nebraska-Lincoln
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2131783&HistoricalAwards=false
• 关键词: Mechanisms; Small; RNA; Mediated; Silencing

RNA interference (RNAi) is a cellular process that regulates gene expression and acts as a defense system in plants and animals. This pathway relies on small RNAs (sRNAs) to identify targets, which are then suppressed by a variety of molecular mechanisms. Microalgae are an important and sustainable feedstock for producing diverse biomaterials and nutraceuticals, but improving their productivity is constrained by limited knowledge of how they control gene expression and metabolism. This project will contribute to our understanding of the role of sRNAs in microalgae and to the development of RNAi tools for practical applications (such as RNAi-based crop protection strategies for algal mass culturing). The research will also contribute to the training of undergraduate and graduate students and one postdoctoral fellow. Recruitment of students from underrepresented groups in the sciences, potential first-generation college students, or those that have financial needs will be enabled by participation in several educational programs at the University of Nebraska-Lincoln.Several aspects of sRNA function in eukaryotes, particularly how sRNAs mediate translational repression, remain poorly understood. One key problem is an incomplete knowledge of the components of sRNA-effector complexes and their potential modulation in response to internal developmental programs and/or external cues. This project will provide insight into aspects of sRNA function, particularly the translation repression of polyribosome-associated transcripts. In the alga Chlamydomonas reinhardtii, a homolog of the Drosophila VASA INTRONIC GENE (VIG) is involved in this process. The research will: (i) characterize the interplay between VIG and Argonaute effector complexes in sRNA-mediated translation repression by several proteomic approaches; (ii) isolate additional genes involved in sRNA-mediated translation repression by using an insertional mutagenesis screen; and (iii) characterize the biogenesis and potential function of a novel class of Chlamydomonas sRNAs, similar in length to the metazoan PIWI-interacting RNAs. Since sRNA-mediated repression of polyribosomal mRNAs also occurs in land plants and animals, the results may contribute to our understanding of the evolution (i.e., conservation vs. divergence) of sRNA-mediated epigenetic mechanisms in eukaryotes.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.

RNA 干扰 (RNAi) 是一种调节基因表达并充当动植物防御系统的细胞过程。该途径依赖小 RNA (sRNA) 来识别靶标，然后通过多种分子机制抑制这些靶标。微藻是生产多种生物材料和营养保健品的重要且可持续的原料，但由于对其如何控制基因表达和代谢的了解有限，其生产力的提高受到限制。该项目将有助于我们了解 sRNA 在微藻中的作用，并有助于开发实际应用的 RNAi 工具（例如用于藻类大规模培养的基于 RNAi 的作物保护策略）。该研究还将有助于培养本科生和研究生以及一名博士后研究员。通过参加内布拉斯加大学林肯分校的多个教育项目，可以招募来自科学领域代表性不足的群体、潜在的第一代大学生或有经济需要的学生。 sRNA 在真核生物中发挥作用的几个方面，特别是如何sRNA 介导翻译抑制，但人们对此仍知之甚少。一个关键问题是对 sRNA 效应复合物的组成及其响应内部发育程序和/或外部线索的潜在调节的了解不完整。该项目将深入了解 sRNA 功能的各个方面，特别是多核糖体相关转录本的翻译抑制。在藻类莱茵衣藻中，果蝇 VASA 内电子基因 (VIG) 的同源物参与了这一过程。该研究将：（i）通过几种蛋白质组学方法表征 VIG 和 Argonaute 效应复合物在 sRNA 介导的翻译抑制中的相互作用； (ii) 通过使用插入诱变筛选分离参与 sRNA 介导的翻译抑制的其他基因； (iii) 表征一类新型衣藻 sRNA 的生物发生和潜在功能，其长度与后生动物 PIWI 相互作用 RNA 相似。由于 sRNA 介导的多核糖体 mRNA 抑制也发生在陆地植物和动物中，因此结果可能有助于我们了解真核生物中 sRNA 介导的表观遗传机制的进化（即保守与分化）。该奖项反映了 NSF 的法定使命和通过使用基金会的智力价值和更广泛的影响审查标准进行评估，该项目被认为值得支持。