Molecular and genetic analysis of novel Slicer-dependent miRNA pathways in blood

血液中新型 Slicer 依赖性 miRNA 通路的分子和遗传分析

• 批准号: 9219500
• 负责人: Michael Kharas
• 金额: $ 69.86万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-10 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9219500
• 关键词: Address; Archaea; Basophilic Erythroblast; Biochemical; Biochemical Genetics; Biogenesis; Biological; Biology; Blood; Catalysis; Cells; Cleaved cell; Complex; Data; Defect; Deposition; Development; Disease; Engraftment; Enzymes; Erythroid; Exhibits; Foundations; Generations; Genetic; Genetic Enhancement; Genetic Epistasis; Genetic study; Genomics; Heart; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic System; Homeostasis; In Vitro; Individual; Knock-in Mouse; Knock-out; Knockout Mice; Malignant - descriptor; Malignant Neoplasms; Mammals; Mediating; Methods; MicroRNAs; Molecular Analysis; Molecular Genetics; Mus; Mutant Strains Mice; Natural regeneration; Nucleotides; PTEN gene; Pathway interactions; Perinatal; Phenotype; Play; Positioning Attribute; Post-Transcriptional Regulation; Process; Proteins; RNA; RNA Interference; Reaction; Regulation; Regulator Genes; Reporting; Repression; Role; Series; Severities; Signal Transduction; Slice; Small RNA; Stress; Study models; System; T-Lymphocyte; Testing; Transcript; Virus; Work; base; cohort; crosslinking and immunoprecipitation sequencing; erythroid differentiation; experimental study; functional loss; genetic analysis; genome-wide; in vivo; insight; knockout animal; leukemia; loss of function; man; mutant; novel; overexpression; oxidant stress; response; transcriptome; transcriptome sequencing

PROJECT ABSTRACT Molecular and genetic analysis of novel Slicer-dependent miRNA pathways in blood Most conserved microRNAs (miRNAs) are generated by a biogenesis pathway that deposits them into an Argonaute effector, guiding them to broad regulatory target networks. Amongst the cohort of four mammalian Argonautes, only Ago2 has catalytic ability to cleave transcripts, an enzymatic activity known as "Slicing" that underlies experimental RNA interference. Nevertheless, the endogenous biological usage of mammalian Slicing remains largely mysterious. Our previous and ongoing studies provide the unexpected perspective of multiple Slicing-dependent biogenesis strategies that generate both Dicer-independent and Dicer-dependent erythroid miRNAs. These data strongly support our hypothesis that a dominant usage of Ago2 catalysis is to generate specific conserved miRNAs in the blood system. Our extensive preliminary data are the basis of (1) a series of biochemical and genomic experiments to elucidate a novel Slicing-dependent miRNA biogenesis mechanism, (2) genetic studies of novel knockout animals of erythroid, Slicing-dependent miRNAs in normal development, blood homeostasis and leukemia, and (3) molecular genetic analyses that seek to connect dysregulated processes in Ago2-catalytically defective blood system to specific Slicing- dependent miRNAs. These studies will bring new insights on post-transcriptional control of erythroid development, homeostasis, and blood cancer, as well as pinpoint the functional basis of mammalian RNAi to the generation of erythroid-specific miRNAs.

项目摘要 血液中依赖Slicer的miRNA新途径的分子和遗传分析 大多数保守的microRNAs(MiRNAs)是由生物发生途径产生的 这使他们成为Argavite的效应器，引导他们达到广泛的监管目标 网络。在四种哺乳动物中，只有Ago2有催化作用。 切割转录本的能力，这是一种称为“切片”的酶活动，它是 实验性的RNA干扰。尽管如此，内源生物利用 哺乳动物的切片在很大程度上仍然是个谜。我们之前和正在进行的研究 提供多种依赖切片的生物发生策略的意想不到的视角 可同时产生非依赖和依赖的红系miRNAs。这些 数据有力地支持了我们的假设，即Ago2催化的主要用途是 在血液系统中产生特定的保守miRNAs。我们广泛的初步调查 数据是(1)一系列生化和基因组实验的基础，以阐明 新的依赖切片的miRNA生物发生机制，(2)新的遗传学研究 红系基因敲除动物，依赖切片的miRNAs在正常发育中，血液 动态平衡和白血病，以及(3)寻求联系的分子遗传分析 在Ago2催化的缺陷血液系统中对特定切片的失调过程- 依赖的miRNA。这些研究将为转录后调控带来新的见解 红系发育、动态平衡和血癌的研究，以及精确定位 哺乳动物RNAi对红系特异性miRNAs产生的功能基础。