Movement of RNA between animal cells

RNA在动物细胞之间的运动

• 批准号: 9334884
• 负责人: Antony Merlin Jose
• 金额: $ 28.65万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9334884
• 关键词: Address; Affect; Animals; Biochemical Genetics; Biogenesis; Biological Assay; Biology; Blood; Caenorhabditis elegans; Cell Nucleus; Cells; Defect; Diagnostic; Disease; Distant; Double-Stranded RNA; Embryo; Embryonic Development; Endocytosis; Family; Formulation; Gene Silencing; Gene Targeting; Genes; Genetic Transcription; Goals; Homologous Gene; Human; Integral Membrane Protein; Lead; Light; Mammalian Cell; Mammals; Membrane; MicroRNAs; Modeling; Modification; Movement; Mutation; Neurons; Northern Blotting; Organ; Orthologous Gene; Pathway interactions; Pharmaceutical Preparations; Plants; Population; Process; Production; Protein Tyrosine Kinase; Proteins; RNA; RNA Binding; RNA Interference; RNA Interference Therapy; RNA Transport; Reporter; Role; Signal Transduction; Small RNA; Source; Testing; Therapeutic; Tissues; Work; base; design; ds RNA-Binding Proteins; experimental study; extracellular; genetic approach; human disease; in vitro Assay; in vitro activity; insight; member; mutant; novel; nucleotidyltransferase; overexpression; public health relevance; tool; transcriptome sequencing; uptake

DESCRIPTION (provided by applicant): The use of double-stranded RNA (dsRNA) to trigger RNA interference and thereby silence matching disease genes is being pursued as a therapeutic approach for many human diseases. However, it is difficult to deliver these RNA-based drugs into human cells. In contrast, when dsRNA is introduced into one cell in plants and in some animals such as the worm C. elegans, genes in distant cells are also silenced due to the transport between cells of forms of dsRNA called mobile RNA. This direct transport of RNA between cells represents a novel form of intercellular signaling that was accidentally discovered as part of gene silencing studies in the worm C. elegans and in plants. How RNA that is transcribed in the nucleus is secreted from animal cells to silence a target gene in distant cells s not well understood. Studies in C. elegans identified conserved proteins required for the uptake of mobile RNAs suggesting that similar mechanisms may exist in other animals, including mammals. Intriguingly, populations of extracellular RNAs have been found in human blood and are being pursued as diagnostic tools to assess the disease state of internal organs, which are presumably the source of such RNAs. The goals of this proposal are to use the tools available in C. elegans to uncover how dsRNA is modified to generate mobile RNAs that are secreted from cells and to analyze genes that control the export of such RNAs from cells. Using available mutants, we will take biochemical and genetic approaches to identify modified dsRNAs and the mobile RNAs that result from them. We will use new mutants we have isolated that are defective in the intercellular transport of RNAs to discover additional genes that control this process and analyze them. The completion of these aims can reveal how RNAs are modified for export from animal cells and can begin to assemble the pathway for intercellular signaling by the direct transport of RNA between animal cells. These studies are also relevant to understand the entirely unknown biology of extracellular RNAs, which can become powerful diagnostic tools for diseases of internal organs. Finally, if we understand what modifications to dsRNA and other cellular machinery enable the efficient entry of mobile RNAs into C. elegans cells, we can design similar modified dsRNAs for efficient delivery into human cells for RNA interference-based therapy.

描述（由申请人提供）：使用双链RNA（dsRNA）来触发RNA干扰，从而使匹配的疾病基因沉默，正被用作许多人类疾病的治疗方法。然而，很难将这些基于RNA的药物递送到人类细胞中。相反，当双链RNA被导入植物和某些动物（如蠕虫C.在线虫中，由于称为移动的RNA的dsRNA形式在细胞之间的运输，远处细胞中的基因也被沉默。这种RNA在细胞间的直接运输代表了一种新的细胞间信号传导形式，这是在蠕虫C的基因沉默研究中偶然发现的。elegans线虫and in plants植物.在细胞核中转录的RNA是如何从动物细胞中分泌出来，使远处细胞中的靶基因沉默的，目前还不清楚。在C.线虫鉴定了摄取移动的RNA所需的保守蛋白，这表明类似的机制可能存在于其他动物，包括哺乳动物。有趣的是，在人类血液中发现了细胞外RNA的群体，并被用作评估内脏器官疾病状态的诊断工具，这些器官可能是这些RNA的来源。这个提议的目标是使用C中可用的工具。elegans来揭示dsRNA如何被修饰以产生从细胞分泌的移动的RNA，并分析控制这些RNA从细胞输出的基因。利用现有的突变体，我们将采取生物化学和遗传学的方法来鉴定修饰的dsRNA和由它们产生的移动的RNA。我们将使用我们已经分离出的在RNA的细胞间运输中有缺陷的新突变体来发现控制这一过程的其他基因并对其进行分析。这些目标的完成可以揭示RNA是如何被修饰以从动物细胞输出的，并且可以开始通过RNA在动物细胞之间的直接运输来组装细胞间信号传导的通路。这些研究也与了解细胞外RNA的完全未知的生物学有关，细胞外RNA可以成为内脏疾病的强大诊断工具。最后，如果我们了解了对dsRNA和其他细胞机制的修饰使移动的RNA能够有效地进入C。elegans细胞，我们可以设计类似的修饰的dsRNA，用于有效递送到人类细胞中，用于基于RNA干扰的治疗。