Identification of mRNA decay networks in the Drosophila nervous system

果蝇神经系统中 mRNA 衰减网络的鉴定

• 批准号: 8220864
• 负责人: Michael Cleary
• 金额: $ 22.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, MERCED
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8220864
• 关键词: 4-thiouracil; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Biological; Biological Assay; Biological Process; Cells; Dactinomycin; Data Analyses; Defect; Development; Disease; Drosophila genus; Drosophila melanogaster; Elements; Embryo; Embryonic Nervous System; Equilibrium; Family; Gene Expression; Generations; Genes; Genetic Transcription; Goals; Half-Life; Human; In Vitro; Measurement; Measures; Messenger RNA; Methods; Microarray Analysis; Nervous System Physiology; Nervous system structure; Neurons; Orthologous Gene; Pathway interactions; Pharmaceutical Preparations; Physiologic pulse; Physiological; Population Heterogeneity; Presynaptic Terminals; Process; RNA Decay; RNA Interference; RNA Sequences; RNA Stability; Regulation; Regulatory Element; Reporter; Role; Signal Transduction; Site; Structure; System; Techniques; Testing; Transcript; Uracil phosphoribosyltransferase; Uridine; Validity of Results; Work; base; cell type; computerized tools; genetic regulatory protein; genome-wide; genome-wide analysis; in vivo; knock-down; nerve stem cell; nervous system development; nervous system disorder; neurodevelopment; neurogenesis; new technology; novel; prevent; programs; public health relevance; relating to nervous system; research study; response; uracil analog

DESCRIPTION (provided by applicant): Gene expression levels are determined by the transcription rate of each gene and the decay rate of the resulting mRNA. The balance between these two processes can be altered in response to physiologic conditions, developmental signals, or disease states. Regulation of mRNA decay is particularly important in the nervous system, where the unique structure of neurons requires mRNAs to be selectively stabilized in axon terminals, far from their site of synthesis, and the generation of cellular diversity by neural progenitors requires the programmed decay of mRNAs that regulate proliferation or differentiation. Defective regulation of mRNA decay has been implicated in several human neurological disorders, including Alzheimer's disease and amyotrophic lateral sclerosis. To completely understand the role of mRNA decay in neural development and disease, it will be useful to construct mRNA decay regulatory networks that contain the following information: genome-wide mRNA decay rates in specific cell types under defined conditions, the regulatory proteins that target specific mRNAs for decay, and the mRNA sequence elements that confer coordinate programs of decay. To identify the components of such a network, it is necessary to measure mRNA decay in specific cell types, on a genome-wide scale, under in vivo conditions. Such experiments have historically been difficult, if not impossible, to perform. This project uses a novel technique, known as "TU-tagging", that allows mRNA decay measurements from specific cell types under in vivo conditions. We have recently developed this technique in the model organism, Drosophila melanogaster, and are now well poised to use TU-tagging to identify mRNA decay regulatory networks in the Drosophila nervous system. Our aims are to optimize and validate the accuracy and sensitivity of TU-tagging-based mRNA decay measurements, measure mRNA stability in cells of the Drosophila nervous system, identify the mRNA targets of an mRNA decay regulatory factor encoded by the found in neurons (FNE) gene, and identify cis-regulatory elements among coordinately regulated mRNAs. The specific aims of this work encompass both technical goals and biological goals, reflecting the potential of this project to make significant contributions to the scientific "toolkit" as well as advance the understanding of fundamental biological processes that are relevant to human neural development and disease. PUBLIC HEALTH RELEVANCE: The development and function of the nervous system requires precise control of gene expression and the decay of messenger RNAs (mRNAs) is an essential yet often overlooked control mechanism. Altered regulation of mRNA decay has been implicated in developmental defects and neurological disorders, including Alzheimer's disease and amyotrophic lateral sclerosis. We have developed a novel technology that will allow genome-wide analysis of mRNA decay in neural cells and the construction of comprehensive "blueprints" of mRNA decay pathways that control nervous system development and function.

描述（由申请人提供）： 基因表达水平由每个基因的转录率和所得 mRNA 的衰减率决定。这两个过程之间的平衡可以根据生理条件、发育信号或疾病状态而改变。 mRNA 衰变的调节在神经系统中尤其重要，其中神经元的独特结构要求 mRNA 选择性地稳定在远离其合成位点的轴突末端，而神经祖细胞产生细胞多样性需要调节增殖或分化的 mRNA 的程序性衰变。 mRNA 衰变的调节缺陷与多种人类神经系统疾病有关，包括阿尔茨海默病和肌萎缩侧索硬化症。为了完全理解 mRNA 衰变在神经发育和疾病中的作用，构建包含以下信息的 mRNA 衰变调节网络将很有用：特定细胞类型在特定条件下的全基因组 mRNA 衰变率、针对特定 mRNA 衰变的调节蛋白以及赋予衰变协调程序的 mRNA 序列元件。为了识别这种网络的组成部分，有必要在体内条件下在全基因组范围内测量特定细胞类型中 mRNA 的衰减。从历史上看，这样的实验即使不是不可能，也是很难进行的。 该项目使用了一种称为“TU 标记”的新技术，可以在体内条件下测量特定细胞类型的 mRNA 衰减。我们最近在模型生物体果蝇中开发了这项技术，现在已经准备好使用 TU 标签来识别果蝇神经系统中的 mRNA 衰变调节网络。我们的目标是优化和验证基于 TU 标记的 mRNA 衰减测量的准确性和灵敏度，测量果蝇神经系统细胞中 mRNA 的稳定性，识别由神经元 (FNE) 基因编码的 mRNA 衰减调节因子的 mRNA 靶标，并识别协调调节的 mRNA 中的顺式调节元件。这项工作的具体目标包括技术目标和生物学目标，反映了该项目为科学“工具包”做出重大贡献以及增进对与人类神经发育和疾病相关的基本生物过程的理解的潜力。 公共卫生相关性： 神经系统的发育和功能需要精确控制基因表达，而信使 RNA (mRNA) 的衰变是一种重要但经常被忽视的控制机制。 mRNA 衰变调节的改变与发育缺陷和神经系统疾病有关，包括阿尔茨海默病和肌萎缩侧索硬化症。我们开发了一种新技术，可以对神经细胞中的 mRNA 衰变进行全基因组分析，并构建控制神经系统发育和功能的 mRNA 衰变途径的全面“蓝图”。