Double-stranded RNA-mediated signaling pathway and gene silencing

双链RNA介导的信号通路和基因沉默

• 批准号: 7763908
• 负责人: YI LIU
• 金额: $ 27.98万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7763908
• 关键词: Address; Adoption; Antiviral Agents; Biochemical; Biogenesis; Biological Models; Bloom syndrome protein; Cells; DNA Binding; DNA Damage; DNA Double Strand Break; DNA Repair; DNA biosynthesis; Defense Mechanisms; Development; Double Strand Break Repair; Double-Stranded RNA; Eukaryota; Foundations; Gene Activation; Gene Expression; Gene Expression Regulation; Gene Silencing; Generations; Genes; Genetic; Genetic Screening; Genetic Transcription; Genome Stability; Goals; Homologous Gene; Host Defense; Human; Immune; Impairment; Interferons; Lead; Link; Maintenance; Malignant Neoplasms; Mammals; Mediating; Molds; Names; Natural Immunity; Neurospora; Neurospora crassa; Nucleotides; Organism; Pathway interactions; Pharmacologic Substance; Physiological; Process; Production; Proteins; RNA; RNA Interference; RNA Interference Pathway; RNA-Directed RNA Polymerase; Recombinant DNA; Recruitment Activity; Regulation; Replication-Associated Process; Role; Signal Pathway; Signaling Pathway Gene; Single-Stranded DNA; Small Interfering RNA; Small RNA; System; Technology; Testing; Transcript; Transcriptional Activation; Transcriptional Regulation; Validation; Virus Diseases; base; defense response; helicase; human disease; mutant; novel; novel therapeutic intervention; positional cloning; protein complex; public health relevance; repaired; replication factor A; response; sensor; therapeutic development; viral RNA

DESCRIPTION (provided by applicant): Double-stranded RNA (dsRNA) production in cells is known to activate the RNA interference (RNAi) pathway in almost all eukaryotic organisms and to trigger the transcription-based interferon response in mammals. The filamentous fungus Neurospora crassa, an organism that broadly employs gene silencing in regulation of gene expression, offers a unique and powerful system for understanding the RNAi pathway and its function in eukaryotes. We showed that the Neurospora RNAi pathway, like those in higher eukaryotes, uses small RNAs to mediate posttranscriptional gene silencing. We also showed that dsRNA leads to the transcriptional activation of many genes in Neurospora, including most of the RNAi components, putative antiviral genes, and homologs of the interferon stimulated genes. Furthermore, we discovered a novel type of small RNA that is dramatically induced after DNA damage. In addition, we found that DNA damage results in the production aberrant RNA from the loci where small RNAs are produced. These studies suggested evolutionarily conserved roles of RNAi in gene silencing and defense mechanisms and a novel link between RNAi pathway and DNA repair/replication processes. In Specific Aim 1, we will determine the signaling pathway responsible for dsRNA-induced gene transcription. We will identify the components of this pathway by both forward and reverse genetic approaches. This study will lead to the understanding of a novel dsRNA activated signaling pathway that may be relevant in mammals. In Specific Aim 2, we will determine the biogenesis and function of the DNA damage- induced small RNA. This study will reveal the role and the mechanism of RNAi in DNA repair and in maintaining genome stability. In addition, we will investigate the mechanism of aberrant RNA production after DNA damage. These proposed studies, using a combination of genetic, biochemical and physiological approaches, will have important implications for the understanding of eukaryotic gene silencing and defense responses in general. PUBLIC HEALTH RELEVANCE RNA interference (RNAi) and related pathways regulate gene expression, development, genome stability and defense responses in human. The dsRNA-induced interferon response is an important part of the human innate immunity toward viral infection. The impairment of these pathways can result in developmental abnormality, cancer or immune deficiency. Furthermore, development and adoption of RNAi technologies have been extensively used in pharmaceutical target validation and in therapeutic development. Our goal is to understand the function and regulation of these pathways in human using a simple eukaryotic model system. A better understanding of these pathways will potentially lead to new therapeutic approaches for treating human diseases.

描述（由申请人提供）：已知细胞中的双链RNA（DSRNA）产生可激活几乎所有真核生物中的RNA干扰（RNAI）途径，并在哺乳动物中触发基于转录的干扰素反应。丝状真菌神经孢子虫是一种广泛采用基因沉默的生物体，用于调节基因表达的调节，为理解RNAi途径及其在真核生物中的功能提供了独特而强大的系统。我们表明，像较高的真核生物中的神经孢子虫RNAi途径使用小的RNA来介导转录后基因沉默。我们还表明，dsRNA导致神经孢子中许多基因的转录激活，包括大多数RNAi成分，假定的抗病毒基因和干扰素刺激基因的同源物。此外，我们发现了一种新型的小RNA，该RNA在DNA损伤后急剧诱导。此外，我们发现DNA损伤导致产生小RNA的基因座的产生异常RNA。这些研究表明，RNAi在基因沉默和防御机制中的进化保守作用，以及RNAi途径与DNA修复/复制过程之间的新联系。在特定目标1中，我们将确定负责DSRNA诱导基因转录的信号传导途径。我们将通过前进和反向遗传方法确定该途径的组成部分。这项研究将导致对可能与哺乳动物有关的新型DSRNA激活信号通路的理解。在特定的目标2中，我们将确定DNA损伤诱导的小RNA的生物发生和功能。这项研究将揭示RNAi在DNA修复和维持基因组稳定性中的作用和机制。此外，我们将研究DNA损伤后异常RNA产生的机制。这些提出的研究结合了遗传，生化和生理方法的结合，将对理解真核基因沉默和一般的防御反应具有重要意义。 公共卫生相关性RNA干扰（RNAI）和相关途径调节人类基因表达，发展，基因组稳定性和防御反应。 DSRNA诱导的干扰素反应是人类对病毒感染的先天免疫的重要组成部分。这些途径的损害会导致发育异常，癌症或免疫缺陷。此外，RNAi技术的开发和采用已被广泛用于药物靶向验证和治疗开发中。我们的目标是使用简单的真核模型系统了解人类中这些途径的功能和调节。更好地了解这些途径将有可能导致治疗人类疾病的新治疗方法。