TRANSGENIC EXPRESSION ANALYSIS OF MIRNA-144

miRNA-144 的转基因表达分析

• 批准号: 8168363
• 负责人: MICHAEL D WESTON
• 金额: $ 28.55万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168363
• 关键词: 3' Untranslated Regions; A Mouse; Affect; Binding; Biological; Biological Assay; Communities; Computer Retrieval of Information on Scientific Projects Database; Development; Disease; Exhibits; Family; Funding; Gene Targeting; Grant; Hair Cells; Hearing; Histologic; Homeostasis; Human; Institution; Knowledge; Labyrinth; Luciferases; Mediating; MicroRNAs; Microarray Analysis; Modality; Mus; Organ of Corti; Pathway interactions; Phenotype; Physiological; Presbycusis; Production; Proteins; RNA; RNA Interference; Regulation; Reporter; Research; Research Personnel; Resources; Sensory Hair; Site; Source; Supporting Cell; Therapeutic; Time; Tissues; Transgenic Organisms; United States National Institutes of Health; Untranslated RNA; hearing impairment; mouse model; notch protein; novel; spatiotemporal; transcription factor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. MicroRNAs (miRNAs) are small noncoding RNAs that regulate protein production of target mRNAs through recognition and binding of partially complementary sites within 3' untranslated regions. The miRNA-183 family (miR-183fam) are a trio of evolutionary conserved neurosensory miRNAs whose coordinated spatiotemporal expression in the mouse inner ear suggests they are long term markers essential to sensory hair cell identity and function. A mouse model of miR-183fam misexpression has been generated which exhibits an age-related hearing loss phenotype. The hypothesis is that the timing and strength of miR-183fam abundance is critical to the normal development and homeostasis of mouse inner ear sensory hair cells and supporting cells through RNAi mediated regulation of downstream target genes. This proposal aims to: 1) complete histologic and physiologic assessments of FVB/N Tg(GFAP-183-96-182) homozygous mice to correlate its inner ear phenotype with miR-183fam misexpression in the organ of Corti and identify the earliest time and type of deficit which causes the age-related hearing loss; 2) validate predicted miRNA-183fam regulation of notch receptor Jag1 and the downstream transcription factor Sox2 with heterologous luciferase reporter assays and through assessments of RNA and protein levels in affected transgenic tissues; 3) identify primary biological pathways and candidate target genes changed by miR-183fam misexpression through organ of Corti Affymetrix microarray analysis. Direct effects on inner ear function through the modulation of specific miRNAs establishes a paradigm for novel miRNA mediated therapeutics to treat some form of human hearing loss. Elucidation of the mechanism Tg-miRNA mediated hair cell loss should yield modalities influencing organ of Corti homeostasis, which may be of therapeutic benefit to treat hearing loss disease. Completion these aims will serve as paradigm for elucidating miRNA function in general. Knowledge gained from this study will significantly influence the miRNA and hearing research communities by demonstrating miRNAs as crucial and potent factors in the development and homeostasis of the inner ear.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 microRNA（miRNAs）是一类非编码小RNA，通过识别和结合3'端非翻译区的部分互补位点来调控靶mRNA的蛋白质合成。miRNA-183家族（miR-183 fam）是进化上保守的三种感觉神经miRNA，其在小鼠内耳中的协调时空表达表明它们是感觉毛细胞身份和功能所必需的长期标记物。已经产生了miR-183 fam错误表达的小鼠模型，其表现出年龄相关的听力损失表型。该假说认为，miR-183 fam丰度的时间和强度通过RNAi介导的下游靶基因调节对小鼠内耳感觉毛细胞和支持细胞的正常发育和稳态至关重要。该提案旨在：1）FVB/N Tg的完整组织学和生理学评估（GFAP-183-96-182）纯合小鼠的内耳表型与Corti器官中miR-183 fam的错误表达相关联，并鉴定导致年龄相关性听力损失的最早缺陷时间和类型; 2）验证预测的miRNA-用异源荧光素酶报告基因测定和通过RNA和RNA水平评估notch受体Jag 1和下游转录因子Sox 2的183 fam调节受影响的转基因组织中的蛋白质水平; 3）通过Corti Affyssin微阵列分析鉴定由miR-183 fam错误表达改变的主要生物学途径和候选靶基因。通过特定miRNA的调节对内耳功能的直接作用建立了用于治疗某种形式的人类听力损失的新型miRNA介导的疗法的范例。阐明Tg-miRNA介导的毛细胞损失的机制将产生影响Corti器官稳态的模式，这可能对治疗听力损失疾病具有治疗益处。这些目标的完成将作为阐明miRNA功能的范例。从这项研究中获得的知识将通过证明miRNA是内耳发育和动态平衡的关键和有效因素，对miRNA和听力研究界产生重大影响。