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

分子生物学的核心法则认为，遗传密码存在于DNA（脱氧核糖核酸）中，DNA（脱氧核糖核酸）产生RNA（核糖核酸）转录本，这些转录本随后被翻译成最终影响生物体功能的蛋白质。在过去的十年里，RNA的作用已经大大超越了中间分子的作用，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所起的作用，包括发育和疾病进展。该项目的目标是更好地了解mirna在非模式系统中的作用，特别是来自南大洋的鱼类，同时，这将使人们了解这些只在南极洲发现的鱼类对细胞压力的反应能力。除了对海洋变化（更高的温度、更低的pH值、不断变化的盐度）的担忧之外，南大洋内的捕鱼活动也显著增加。因此，更清楚地了解极地鱼类种群的易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。该项目还将为初级本科院校（PUI）的学生提供前沿研究机会，帮助学生建立在日益技术化的研究领域取得成功所需的工具。该项目将整合几种分子方法来功能表征微rna （miRNAs）及其在调节南极鱼细胞应激反应中的作用。先前的研究工作已经提供了证据，支持这些鱼的一部分细胞应激反应失去了调节控制。该团队将对这些鱼的mirna表达进行转录组分析，并将这些表达模式与高温下应激反应基因mRNA（信使RNA）丰度的变化联系起来。此外，PI？我们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来对推测的mirna进行功能表征。这些功能分析将有助于确定miRNA是否对一组辅助蛋白质折叠的关键分子伴侣的RNA转录物的丰度进行调控。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关键见解。尽管在自然界中存在高风险（这项工作很少在非模式物种中进行），但该项目将为研究基因表达变化和一系列生理变化之间的调节机制提供一个前所未有的窗口，从整个生物体到细胞，在单个非模式鱼中进行测量。

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.