Recognition of Cellular Targets by single and double-stranded nucleic acids

单链和双链核酸对细胞靶标的识别

• 批准号: 9071164
• 负责人: David R Corey
• 金额: $ 6.18万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9071164
• 关键词: Affect; Alleles; C9ORF72; Cell Nucleus; Cells; Cellular biology; Chemistry; Cytoplasm; Data; Disease; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Goals; Laboratories; Lead; Mammalian Cell; Mass Spectrum Analysis; Mediating; Messenger RNA; Methods; Molecular; Nuclear; Nuclear RNA; Nucleic Acids; Pathway interactions; Property; Proteins; Publishing; RNA; RNA Interference; RNA Splicing; RNA-Protein Interaction; Reporting; Somatic Cell; Structure; Telomerase; Translations; Untranslated RNA; Validation; Work; cellular targeting; disorder control; driving force; drug development; frataxin; insight; mutant; novel; protein expression; public health relevance; telomere; transcriptome sequencing

 DESCRIPTION (provided by applicant): My laboratory investigates the properties of natural and chemically modified nucleic acids. Working at the interface of cell biology and nucleic acid chemistry we have modified cellular activities including transcription, translation, allele-selectie inhibition of protein expression, splicing, and telomerase-mediated elongation of telomeres. For 2016-2020 our central goal is to examine the recognition of cellular RNA with a primary focus on nuclear RNA. We will apply that understanding to methods for controlling gene expression and new insights into natural pathways of gene regulation. Much of the genome is transcribed into RNAs that do not encode proteins. The function of many noncoding RNAs is unclear, as is the mechanism for how they might affect gene expression. Published reports offer little insight into the molecular details for how a specific noncoding RNA affects expression of a given gene. In the absence of this information, validation of proposed effects becomes problematic and the predictive power for studying novel genes is limited. This lack of guiding mechanistic principles is a major obstacle to progress. RNAi provides a potential mechanism for recognizing specific nuclear RNA species. While RNAi is well-known as a driving force for recognition of mRNA in the cytoplasm of mammalian cells, its potential to drive recognition in the somatic cell nuclei has been unclear. We propose to define the scope and mechanism of mammalian nuclear RNAi and to apply it to novel disease targets. Objective 1. Understand nuclear RNAi. We will characterize the protein and RNA interactions involved in nuclear RNAi. RNAseq will be used to identify RNA targets for functional validation. Mass spectrometry will be used to determine protein partners. We will explore the mechanism of nuclear RNAi and obtain insights into the how nuclear RNAi regulates gene expression in mammalian cells. These data will reveal how argonaute proteins and other RNAi factors function in the nucleus to control gene expression. Objective 2. Novel targets for nucleic acids inside cells. We will also expand recognition to novel nucleic acids targets including the expanded intronic repeats within the mutant frataxin and C9orf72 genes. These data will further develop our understanding of the mechanism of nuclear RNAi and show how nuclear RNAi can be applied to the control of disease gene expression. Compounds that control frataxin protein expression or disrupt structures formed by the expanded repeat within intronic C9orf72 would be lead compounds for drug development.

 描述（由申请人提供）：我的实验室研究天然和化学修饰的核酸的特性，在细胞生物学和核酸化学的界面上工作，我们改变了细胞活性，包括转录、翻译、蛋白质表达的等位基因选择性抑制、剪接。 ，以及端粒酶介导的端粒延长 2016-2020 年，我们的中心目标是检查细胞 RNA 的识别，主要关注核 RNA。对控制基因表达的方法的了解以及对基因调控自然途径的新见解许多非编码RNA的功能以及它们如何影响基因的机制尚不清楚。已发表的报告对特定非编码RNA如何影响给定基因的表达的分子细节知之甚少，在缺乏这些信息的情况下，对所提出的影响的验证就变得有问题，并且研究新基因的预测能力是有限的。指导机械原理是RNAi 提供了识别特定核 RNA 种类的潜在机制，虽然众所周知，RNAi 是识别哺乳动物细胞质中 mRNA 的驱动力，但它在体细胞核中驱动识别的潜力却很大。 我们建议定义哺乳动物核 RNAi 的范围和机制，并将其应用于新的疾病靶标。我们将描述核 RNAi 中涉及的蛋白质和 RNA 相互作用。用于功能验证的 RNA 靶标。我们将探索核 RNAi 的机制，并深入了解核 RNAi 如何调节哺乳动物细胞中的基因表达。 RNAi 因子在细胞核中发挥作用以控制基因表达。 目标 2. 细胞内核酸的新靶标 我们还将扩大对新核酸靶标的识别，包括突变体 frataxin 和 C9orf72 基因中的扩展内含子重复序列。我们对核 RNAi 机制的理解，并展示了如何将核 RNAi 应用于控制疾病基因表达。控制 frataxin 蛋白表达或破坏内含子内扩展重复形成的结构的化合物。 C9orf72 将成为药物开发的先导化合物。