EAGER: RUI: Functional Characterization of Micro RNAs and their Role in the Cellular Stress Response of Polar Fish

EAGER：RUI：微小 RNA 的功能表征及其在极地鱼细胞应激反应中的作用

• 批准号: 1522865
• 负责人: Sean Place
• 金额: $ 10万
• 依托单位: Sonoma State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-15 至 2017-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1522865&HistoricalAwards=false
• 关键词: EAGER; RUI; Functional; Characterization; Micro

The central dogma of molecular biology states that the genetic code is housed in DNA (DeoxyriboNucleic Acid), from which RNA (RiboNucleic Acid) transcripts are produced that are then translated into proteins that ultimately affect the function of an organism. Within the last decade, the role of RNA has dramatically expanded beyond that of an intermediary molecule, and small RNA (including microRNA, or miRNA) research has rapidly expanded, shedding light on the role non-coding RNAs play in many aspects of biology, including development and disease progression. The goal of this project is to better understand the roles of miRNAs in non-model systems, specifically fishes from the Southern Ocean, which will, at the same time, generate insight into the capacity of these fishes that are only found in Antarctica to respond to cellular stress. In addition to concerns about changing oceans (higher temperatures, lower pH, changing salinities), fishing within the Southern Ocean has increased significantly. Thus, a more clear understanding of the susceptibility of polar fish populations will lead to a greater understanding of how to better approach management of these unique marine ecosystems. This project will also provide cutting edge research opportunities for students at a Primarily Undergraduate Institution (PUI), helping the students build the tools necessary to succeed in an increasingly technical research field.This project will integrate several molecular approaches to functionally characterize microRNAs (miRNAs) and the role they play in regulating the cellular stress response in notothenioid fish. Previous research efforts have provided evidence to support the loss of regulatory control for a portion of the cellular stress response in these fish. The team will undertake a transcriptome wide analysis of the expression of miRNAs in these fish and correlate those expression patterns with changes in mRNA (messenger RNA) abundance of stress response genes under elevated temperatures. Furthermore, the PI?s lab will perform functional characterization of the putative miRNAs by over-expression of these short, non-coding RNAs in cultured fish cells. These functional analyses will help determine if the miRNA displays regulatory control over the abundance of RNA transcripts for a key set of molecular chaperones that assist with protein folding. Ultimately, these studies will shed new light on the possible interruption of the classical heat shock response in these fish and provide critical insight into their capacity to adapt to the expected changes in the Southern Ocean. Although high risk in nature, (this work has rarely been performed in non-model species) this project will provide an unprecedented window into the regulatory mechanisms that link changes in gene expression and a suite of physiological changes, measured from the whole organism to the cell, in an individual, non-model fish.

分子生物学的中心法则指出，遗传密码位于DNA（脱氧核糖核酸）中，从中产生RNA（核糖核酸）转录本，然后翻译成最终影响生物体功能的蛋白质。在过去的十年中，RNA的作用已经大大扩展到中间分子之外，小RNA（包括microRNA或miRNA）的研究也迅速扩大，揭示了非编码RNA在生物学的许多方面发挥的作用，包括发育和疾病进展。该项目的目标是更好地了解miRNAs在非模型系统中的作用，特别是来自南大洋的鱼类，这将同时深入了解这些仅在南极洲发现的鱼类对细胞应激的反应能力。除了对海洋变化（温度升高、pH值降低、盐度变化）的关切外，南大洋的捕捞活动也大幅增加。因此，更清楚地了解极地鱼类种群的敏感性将有助于更好地了解如何更好地管理这些独特的海洋生态系统。该项目还将为一所私立大学本科院校（PUI）的学生提供前沿研究机会，帮助学生构建必要的工具，以便在日益技术化的研究领域取得成功。该项目将整合几种分子方法来表征microRNA（miRNAs）的功能及其在调节脊索鱼细胞应激反应中的作用。以前的研究工作提供了证据，支持这些鱼的细胞应激反应的一部分的监管控制的损失。该团队将对这些鱼类中miRNA的表达进行转录组范围的分析，并将这些表达模式与高温下应激反应基因mRNA（信使RNA）丰度的变化相关联。此外，PI？的实验室将通过在培养的鱼细胞中过表达这些短的非编码RNA来对假定的miRNA进行功能表征。这些功能分析将有助于确定miRNA是否对一组关键的分子伴侣的RNA转录物的丰度进行调控，这些分子伴侣有助于蛋白质折叠。最终，这些研究将为这些鱼类经典热休克反应的可能中断提供新的线索，并为它们适应南大洋预期变化的能力提供重要的见解。虽然在自然界中的高风险，（这项工作很少在非模式物种中进行），该项目将提供一个前所未有的窗口，以调节机制，连接基因表达的变化和一套生理变化，从整个生物体到细胞，在一个单独的，非模式鱼。