Transgenerational gene silencing by extracellular RNA

细胞外RNA的跨代基因沉默

• 批准号: 9894814
• 负责人: Antony Merlin Jose
• 金额: $ 31.41万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-16 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9894814
• 关键词: Affect; Animals; Award; Base Pairing; Biological; Biology; Blood Circulation; C. elegans genome; Caenorhabditis elegans; Cells; Chemicals; Chromatin; Cytosol; Defect; Development; Diabetes Mellitus; Diet; Disease; Double-Stranded RNA; Early Diagnosis; Epigenetic Process; Etiology; Gene Activation; Gene Expression; Gene Expression Regulation; Gene Proteins; Gene Silencing; Generations; Genes; Genetic; Genetic Epistasis; Genetic Screening; Genetic Transcription; Genome Mappings; Germ Cells; Goals; Grant; Guide RNA; Health; Human; Inherited; Length; Longevity; Maintenance; Mammals; Metabolism; Modeling; Modification; Molecular; Mus; Mutate; Nematoda; Neurons; Northern Blotting; Obesity; Pathway interactions; Phenotype; Predisposition; Proteins; RNA; Regulator Genes; Reporter; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Small RNA; Somatic Cell; Starvation; Sterility; Stress; Testing; Tissues; Transgenes; Untranslated RNA; Variant; Work; base; causal variant; epidemiology study; epigenome editing; experience; experimental study; extracellular; genetic approach; genome editing; genome sequencing; histone modification; insight; mRNA sequencing; mortality; mutant; next generation; response; transcriptome sequencing; transmission process; uptake; whole genome

Project Summary Recent epidemiological studies suggest that human health can be influenced by ancestral experience. Ancestral diet and stress have been identified as possible modifiers of health in descendants. However, the mechanisms by which the ancestral experience is transmitted to descendants as gene regulatory information to cause effects is not understood. While RNA and epigenetic modifications have been proposed to correlate with ancestral experience, the complexity of mammalian biology has presented a challenge to establish causal mechanisms. In particular, the molecular agent(s) that transmit the information from one generation to the next and the machinery that promotes changes in gene expression that can be stable for many generations are not well understood. Identifying such markers of ancestral influence can enable the early diagnosis of susceptibility to diseases such as obesity and diabetes. We recently found that RNAs exported from neurons in response to double-stranded RNA (dsRNA) expression can enter germ cells in the nematode C. elegans and silence matching genes in descendants. Intriguingly, the persistence of this silencing depends on as yet unknown features of the gene. Because extracellular RNAs are also found in human circulation, they are candidates for similar transport to the next generation and for triggering subsequent transgenerational effects. In support of this idea, three key aspects appear to be conserved in mammals including humans: use of a dsRNA-selective importer for uptake of extracellular dsRNA, use of Argonaute proteins for gene silencing, and use of histone modifications for stable propagation of the regulatory state of a gene. The goal of this proposal is to understand mechanisms that regulate the ability of extracellular RNA in ancestors to affect genes of matching sequence in descendants. We will determine the features of extracellular RNA that enable it to trigger transgenerational gene silencing using cell biological and genetic approaches. We will determine the factors that make a gene susceptible to transgenerational changes in gene regulation using forward genetic, genome editing, and epigenome editing approaches. Finally, we will identify and validate loci in the C. elegans genome that are susceptible to ancestral influences using insights that we have gained from the silencing of a model gene by dsRNA exported from ancestral neurons. These studies will provide a clear picture of mechanisms that can promote or inhibit gene regulation by inherited RNAs in an animal, which are likely to be extendable to other animals including mammals.

项目摘要 最近的流行病学研究表明，人类健康可能受到祖先经历的影响。 祖先的饮食和压力已被确定为后代健康的可能修饰剂。但 祖先的经验作为基因调节信息传递给后代的机制 造成的影响是不被理解的。虽然RNA和表观遗传修饰被认为与 根据祖先的经验，哺乳动物生物学的复杂性对建立因果关系提出了挑战。 机制等特别是，将信息从一代传递到下一代的分子代理 而促进基因表达变化的机制可以稳定许多代， 很好理解。识别这些受祖先影响的标志物可以使易感性的早期诊断成为可能 肥胖症和糖尿病等疾病。我们最近发现，神经元在响应于 双链RNA（dsRNA）表达可进入线虫C.优雅与沉默 与后代的基因相匹配有趣的是，这种沉默的持久性取决于迄今未知的 基因的特征。由于细胞外RNA也存在于人体循环中，因此它们是 类似的运输到下一代，并引发随后的跨代效应。支持 根据这一想法，包括人类在内的哺乳动物中似乎保留了三个关键方面：使用选择性双链RNA 用于摄取细胞外dsRNA导入剂、Argonaute蛋白用于基因沉默的用途和组蛋白的用途 用于基因的调节状态的稳定繁殖的修饰。本提案的目的是了解 调节祖先细胞外RNA影响匹配序列基因的能力的机制， 后裔我们将确定细胞外RNA的特征，使其能够触发跨代 使用细胞生物学和遗传学方法进行基因沉默。我们将确定一个基因 易受使用正向遗传、基因组编辑的基因调控的代际变化的影响， 表观基因组编辑方法。最后，我们将确定和验证基因座在C。线虫的基因组 利用我们从一个模型基因的沉默中获得的见解， 从祖先神经元输出的dsRNA。这些研究将提供一个清晰的机制， 促进或抑制动物中遗传RNA的基因调控，这些RNA可能会延伸到其他动物。 动物，包括哺乳动物。