RNA Mediated Genetic Interference in C. elegans

RNA 介导的线虫遗传干扰

• 批准号: 10610362
• 负责人: CRAIG C MELLO
• 金额: $ 36.85万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10610362
• 关键词: Animals; Biochemical; Biomedical Research; Caenorhabditis elegans; Cellular biology; Chromatin; Development; Developmental Biology; Double-Stranded RNA; Equipment and supply inventories; Fire - disasters; Gene Expression; Gene Silencing; Generations; Genes; Genetic; Genetic Screening; Genetic Transcription; Genome; Guide RNA; Human; Human Biology; Human Genome; Licensing; Maintenance; Mediating; Messenger RNA; Nematoda; Nucleotides; Pathway interactions; Pharmaceutical Preparations; Play; Protein Family; Proteins; Proteomics; RNA; RNA Interference; Regulator Genes; Ribonuclease H; Role; Scanning; Signal Transduction; Small RNA; System; Technology; Therapeutic; clinical application; clinical development; cofactor; deep sequencing; egg; gene function; genome editing; genome-wide; human disease; insight; recruit; sperm cell; stem cells; tool; transgenerational epigenetic inheritance

RNA interference (RNAi) was originally described as a gene silencing mechanism triggered by the experimental introduction of double stranded (ds)RNA into the nematode C. elegans (Fire et al., 1998). The term RNAi now refers to a diverse set of gene-regulatory mechanisms that share common features, including the involvement of short 21- to 30-nucleotide (nt) RNAs and protein cofactors of the Argonaute (RNase H-related) protein family. As an experimental tool, RN Ai is of broad relevance to basic biomedical research, and RN Ai therapeutics are now approved and under development for several clinical applications. RNAi-related mechanisms function in conserved generegulato1y pathways that are of basic and fundamental importance to human cellular and developmental biology. Remarkably, RNAi-related mechanisms in C.elegans keep inventory of all mRNAs and license gene expression in the germline, passing this information via the egg and sperm from one generation to the next. Distinct Argonaute pathways function in the transgenerational inheritance of small-RNA signals that constitute the CSR-1 "self'/protective (RNAa pathway) and the WAGO "non-self'/silencing (RNAe pathway). A third Argonaute pathway, the PRG-1/Piwi pathway scans germline mRNA for foreign sequences. Aim 1 will explore how these Argonaute pathways identify their targets and recruit downstream factors. Aim 2 will investigate how these Argonaute pathways interact to mediate genome-wide transcriptional surveillance. And finally, Aim 3 will use genetic screens to identify new genes required for RNAi and the related RNAe, RNAa, and newly identified "intronless-silencing" pathways. The ability to combine classical genetics with technology, including deep-sequencing, facile CRlSPR-mediated genome editing, and proteomics, make C. elegans an ideal system for these studies. The mechanisms and protein families that mediate RNAi are highly conserved in animals and therefore insights from the proposed studies will be directly relevant to human biology and disease. These studies will shed light on an ancient gene-regulatory mechanism whose correlates in humans are likely to play important conserved roles in the protection of the human genome and the maintenance of stem cells.

RNA干扰（RNAi）最初被描述为由实验性的干扰所触发的基因沉默机制。 将双链（ds）RNA引入线虫C.秀丽隐杆线虫（Fire等人，1998年）。RNAi这个术语现在指的是 一套多样化的基因调控机制，具有共同的特点，包括参与短21- 30-Argonaute（RNase H相关）蛋白家族的核苷酸（nt）RNA和蛋白辅因子。作为 实验工具，RN Ai与基础生物医学研究具有广泛的相关性，RN Ai疗法现已获得批准 并且正在开发用于几种临床应用。RNAi相关机制在保守基因调控中的作用 这些途径对人类细胞和发育生物学具有基本和根本的重要性。 值得注意的是，秀丽隐杆线虫中的RNAi相关机制保留了所有mRNA的清单并许可基因在细胞中表达 生殖细胞，通过卵子和精子将这些信息从一代传递到下一代。独特的Argonaute 这些途径在构成CSR-1的小RNA信号的跨代遗传中起作用 “自身”/保护性（RNAa途径）和WAGO“非自身”/沉默（RNAe途径）。第三条阿尔戈诺特通道， PRG-1/Piwi途径扫描生殖系mRNA中的外源序列。Aim 1将探索这些Argonaute 途径确定其目标并招募下游因子。Aim 2将研究这些Argonaute通路如何 相互作用以介导全基因组转录监视。最后，Aim 3将使用基因筛选来识别 RNAi所需的新基因以及相关的RNAe、RNAa和新鉴定的“无内含子沉默”途径。 将联合收割机经典遗传学与技术相结合的能力，包括深度测序、容易的CRISPR介导的 基因组编辑和蛋白质组学使C. elegans是这些研究的理想系统。机制和蛋白质 介导RNAi的家族在动物中高度保守，因此， 与人类生物学和疾病直接相关。这些研究将揭示一种古老的基因调控机制 其在人类中的相关物可能在保护人类基因组中发挥重要的保守作用， 干细胞的维持。