MicroRNA expression in mouse ear development

小鼠耳发育中的 MicroRNA 表达

• 批准号: 7275822
• 负责人: MICHAEL D WESTON
• 金额: $ 5.13万
• 依托单位: CREIGHTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7275822
• 关键词: 3' Untranslated Regions; Affect; Binding; Biological; Biological Assay; Biology; CDKN1A gene; Cell Cycle; Cell Differentiation process; Cell Line; Cells; Data; Development; Ear; Ectopic Expression; Embryo; Epithelial; Epithelium; Etiology; Future; Gene Expression Profile; Gene Expression Regulation; Gene Silencing; Gene Targeting; Genes; Glial Fibrillary Acidic Protein; Goals; Grant; Hair; Hair Cells; Hearing problem; Hematopoiesis; Homeostasis; In Situ Hybridization; In Vitro; Intervention; Labyrinth; Ligands; Limb Development; Luciferases; Mediating; Methodology; MicroRNAs; Morphogenesis; Mus; Neurons; Participant; Pathway interactions; Pattern; Play; Probability; Production; Proteins; RNA Interference; Reagent; Regulation; Reporter; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Sensory; Sensory Hair; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Staging; Stem cells; Supporting Cell; System; Testing; Therapeutic; Time; Training; Transgenes; Transgenic Mice; Transgenic Organisms; Translational Repression; Translations; Untranslated RNA; Untranslated Regions; Up-Regulation; angiogenesis; base; design; hearing impairment; in vivo; latent nuclear antigen; lipid biosynthesis; mammalian genome; mouse model; myogenesis; neurogenesis; novel; novel therapeutics; oncoprotein p21; otoconia; postnatal; precursor cell; programs; promoter; receptor; research study; tool; transcription factor

DESCRIPTION (provided by applicant): MicroRNAs are small noncoding RNAs that regulate protein production of target mRNAs through recognition and binding of partially complementary sites located within the 3' untranslated region. The long-term goals of this research are: 1) identify miRNAs critical to inner ear development and homeostasis; 2) establish the degree to which miRNAs dictate inner ear cell fate establishment and homeostasis; 3) identify miRNA directed cellular pathways that intersect hearing disease etiology and implicate signaling pathways and/or novel avenues for potential theraputic intervention. Preliminary data are presented that show the expression of a trio of miRNAs that are significantly and specifically increased concomitant with mouse inner ear maturation. The hypothesis is that the upregulation of miRs 96, 182 and 183 is required for normal maturation and/or homeostasis through its contribution to downstream target gene regulation. Independent miRNA target gene prediction algorthims identify several potential target mRNAs known to be active participants in inner ear biology, including several transcription factors, signaling ligands and receptors. The specific aims of this revised application are: 1) establish the embryonic pattern of miR 96, 182, and 183 expression in the mouse ear; 2) Determine if miRs 96, 182 and 183 inhibit expression of predicted target genes and affect the expression of hair cells marker genes using cell lines from early embryonic mouse inner ear and 3) begin to generate a novel transgenic mouse where endogenous levels of these miRNAs are missexpressed in either the entire ear or supporting cells to study the effects of miR 96,182 and 183 dysregulation in vivo. Fulfillment of the stated objectives will provide novel reagents and a proof of principle that modulation of specific miRNA expression leads to direct effects on inner ear function, thus establishing a paradigm for novel miRNA mediated therapeutics.

描述（由申请人提供）：MicroRNA是小的非编码RNA，其通过识别和结合位于3'非翻译区内的部分互补位点来调节靶mRNA的蛋白质产生。本研究的长期目标是：1）鉴定对内耳发育和稳态至关重要的miRNA; 2）确定miRNA决定内耳细胞命运建立和稳态的程度; 3）鉴定与听力疾病病因学交叉的miRNA指导的细胞通路，并涉及信号通路和/或潜在治疗干预的新途径。初步数据显示，伴随小鼠内耳成熟，三种miRNA的表达显著且特异性增加。该假说是miR 96、182和183的上调通过其对下游靶基因调控的贡献是正常成熟和/或稳态所需的。独立的miRNA靶基因预测算法识别了几种潜在的靶mRNA，已知它们是内耳生物学中的活性参与者，包括几种转录因子、信号配体和受体。该修订申请的具体目标是：1）建立小鼠耳中miR 96、182和183表达的胚胎模式; 2）确定miR 96，182和183使用来自早期胚胎小鼠内耳的细胞系抑制预测的靶基因的表达并影响毛细胞标志物基因的表达，以及3）开始产生一种新的转基因小鼠，其中这些miRNAs的内源性水平在整个耳朵或支持细胞中错误表达，以研究体内miR 96、182和183失调的影响。所述目标的实现将提供新的试剂和原理证明，即特异性miRNA表达的调节导致对内耳功能的直接影响，从而建立新的miRNA介导的治疗的范例。