Genetic Analysis of Conserved microRNAs in C. elegans

线虫中保守 microRNA 的遗传分析

• 批准号: 7456857
• 负责人: Allison Lynn Abbott
• 金额: $ 22.35万
• 依托单位: MARQUETTE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7456857
• 关键词: Accounting; Alleles; Animal Model; Animals; Area; Biological; Biological Assay; Biological Models; Biological Process; C. elegans genome; Caenorhabditis elegans; Cell Differentiation process; Cell physiology; Cells; Collection; Colorectal; Development; EGF Signaling Pathway; Embryo; Erinaceidae; Fibroblast Growth Factor; Fishes; Functional RNA; Gene Expression; Genes; Genetic; Goals; Human; Individual; Knowledge; Life; Lung; Malignant Neoplasms; Mammals; Mediating; Messenger RNA; MicroRNAs; Molecular; Mus; Mutation; Numbers; Pathway interactions; Pattern; Phenotype; Post-Transcriptional Regulation; Process; Public Health; RNA Interference; Regulator Genes; Regulatory Pathway; Relative (related person); Research; Resources; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Study models; Testing; Transcriptional Regulation; Work; carcinogenesis; combinatorial; feeding; genetic analysis; human disease; malignant breast neoplasm; mutant; notch protein; protein expression; response; transcription factor

DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) are small, non-coding RNAs that are indispensable regulators of gene expression. Studies of model organisms have determined that worms, fish, and mice are not able to live without miRNAs. However, little is known about the specific biological pathways and processes regulated by the vast majority of individual miRNAs during animal development. To this end, we have isolated mutant alleles in nearly all miRNA genes in Caenorhabditis elegans. Strikingly, the majority of individual miRNAs are not essential for development or viability. The goal of research in this proposal is to identify a network of genetic interactions between individual miRNAs and other miRNAs, signaling components, and transcription factors. We will focus on those miRNAs that show sequence conservation between worms and mammals. We will use a genetic approach to identify regulatory pathways controlled by miRNAs during C. elegans development. (Aim 1) The hypothesis will be tested that distinct miRNAs function together to regulate cell differentiation, proliferation or survival, possibly by repressing a shared set of target mRNAs. We will analyze the phenotypes of worms with mutations in individual miRNA genes in a sensitized genetic background in which the activity of all miRNAs is reduced. (Aim 2) We will test the hypothesis that miRNAs function to modulate or reinforce signaling pathways, possibly functioning to establish a cell's sensitivity or response to signaling molecules in order to confer robustness of developmental signaling pathways. We will use RNAi to systematically test individual miRNA mutants for genetic interactions with all genes identified as signaling components in the C. elegans genome. (Aim 3) We will test the hypothesis that miRNA-mediated post-transcriptional regulation functions in concert with transcriptional control of shared downstream targets. We will use RNAi to test for functional genetic interactions between individual miRNA genes and all transcription factors in C. elegans. This research will advance our understanding of the principles underlying miRNA action in a cell and will identify functional connections between miRNAs, signaling pathways and transcription factors. Determination of the normal pathways regulated by miRNAs is a critical initial step towards identifying causal relationships between miRNA expression patterns and cancer initiation or progression. Public Health Relevance: MicroRNAs (miRNAs) are small regulatory RNAs that have been implicated in the development or progression of multiple types of human cancers including lung, colorectal, and breast cancer. However, it is not yet clear whether changes in miRNA expression are a cause or consequence of carcinogenesis. In order to elucidate a potential causative role of miRNAs in carcinogenesis, we will use genetic analysis of miRNA mutant C. elegans in order to identify the normal function of miRNAs in cellular processes.

描述（由申请人提供）： microRNA（miRNAs）是一类小的非编码RNA，是基因表达不可或缺的调控因子。对模式生物的研究已经确定，蠕虫、鱼和小鼠不能在没有miRNA的情况下生存。然而，在动物发育过程中，绝大多数个体miRNA调控的特定生物学途径和过程知之甚少。为此，我们在秀丽隐杆线虫几乎所有的miRNA基因中分离出突变等位基因。引人注目的是，大多数单个miRNA对于发育或生存力并不重要。这项研究的目标是确定单个miRNAs与其他miRNAs、信号组分和转录因子之间的遗传相互作用网络。我们将重点关注那些在蠕虫和哺乳动物之间显示序列保守性的miRNA。我们将使用遗传学的方法来确定在C. elegans发育(Aim 1）将测试不同的miRNA一起起作用以调节细胞分化、增殖或存活的假设，可能通过抑制一组共享的靶mRNA。我们将分析在致敏遗传背景中单个miRNA基因突变的蠕虫的表型，其中所有miRNA的活性都降低。(Aim 2）我们将测试这样的假设，即miRNA的功能是调节或加强信号通路，可能起作用以建立细胞对信号分子的敏感性或应答，从而赋予发育信号通路的稳健性。我们将使用RNAi系统地测试单个miRNA突变体与C.线虫基因组(Aim 3）我们将检验miRNA介导的转录后调节与共享下游靶点的转录控制一致发挥作用的假设。我们将使用RNAi来测试单个miRNA基因与C中所有转录因子之间的功能性遗传相互作用。优雅的这项研究将促进我们对细胞中miRNA作用的基本原理的理解，并将确定miRNA，信号通路和转录因子之间的功能联系。确定由miRNA调控的正常途径是确定miRNA表达模式与癌症发生或进展之间因果关系的关键初始步骤。 公共卫生相关性：MicroRNA（miRNAs）是一种小的调控RNA，参与多种人类癌症的发生和发展，包括肺癌、结直肠癌和乳腺癌。然而，目前尚不清楚miRNA表达的变化是致癌的原因还是结果。为了阐明miRNA在肿瘤发生中的潜在致病作用，我们将使用miRNA突变体C的遗传分析。elegans中，以确定在细胞过程中的miRNA的正常功能。