Cis regulatory elements in the inner ear

内耳中的顺式调节元件

• 批准号: 9111307
• 负责人: Olivia Mary Bermingham-McDonogh
• 金额: $ 22.96万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9111307
• 关键词: Adult; Age; Aging; Antibodies; Auditory; BMP4; Binding; Biological Assay; Birds; Brain region; Cell Count; Cell Differentiation process; Cell Line; Cells; ChIP-seq; Chromatin; Cochlear duct; Complex; DNA; DNase I hypersensitive sites sequencing; Defect; Deoxyribonuclease I; Development; Digestion; EP300 gene; Electroporation; Enhancers; Equilibrium; FGF20 gene; Fishes; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genome; Genomics; Hair Cells; Health; Hearing; Human; Hypersensitivity; In Situ Hybridization; Knowledge; Labyrinth; Lead; Maps; Methods; Mus; Mutant Strains Mice; Natural regeneration; Neural Retina; Noise; Nucleic Acid Regulatory Sequences; Organ; Pattern; Pharmaceutical Preparations; Phase; Population; Process; Proteins; Protocols documentation; Regulation; Regulatory Element; Reporting; Research; Role; Sampling; Sensory; Sensory Hair; Signal Transduction; Signaling Molecule; Societies; Staging; Supporting Cell; Techniques; Testing; Time; Tissues; Validation; Vertebrates; Work; cell type; chromatin modification; cognitive development; deafness; ear development; equilibration disorder; functional restoration; gene therapy; hearing impairment; high throughput screening; inner ear diseases; interest; mutant; non-genetic; notch protein; novel; programs; promoter; public health relevance; transcription factor; transcriptome sequencing; transgene expression

 DESCRIPTION (provided by applicant): Hearing loss is a large health issue in the US with approximately 36 million adults reporting some loss. Much of this deafness is caused by loss of the sensory hair cells in the inner ear either from noise or drug damage or just aging. Much research has aimed at replacing these lost cells. Another approach is to restart the developmental program by which these cells develop to begin with. The inner ear is a highly complex tissue, with many different cell types. Understanding the development of this complex organ will ultimately require characterization of gene expression and its regulation in developing and mature inner ear cell types. DNase I hypersensitivity mapping (DNase I-seq) has recently been developed to map enhancers of genes in particular tissues at particular stages of development. In this proposed research we aim to map the enhancers that are used in the inner ear during development. Knowledge of the way genes are regulated in the inner ear may provide us with additional targets for manipulation in our attempts to restore hearing and balance function. This research proposes to completely characterize the enhancers using DNase I hypersensitivity mapping, RNA-seq and using mutant mice that lack sensory domains to determine which are the critical genes. This will be accomplished with the following three specific aims: Aim 1. Mapping the accessible chromatin in the inner ear at three developmental ages. Aim 2. Mapping DNase I hypersensitive regions in a mutant lacking the sensory domain. Aim 3. Characterization of enhancer activity in explant culture.

 听力损失在美国是一个很大的健康问题，大约有3600万成年人报告了一些损失。这种耳聋大部分是由于内耳感觉毛细胞的丧失造成的，无论是噪音还是药物损伤，或者只是衰老。许多研究旨在取代这些丢失的细胞。另一种方法是重新启动这些细胞开始发育的发育程序。内耳是一个高度复杂的组织，有许多不同的细胞类型。了解这个复杂器官的发育最终需要表征基因表达及其在发育和成熟内耳细胞类型中的调节。DNA酶I超敏性作图（DNase I-seq）最近已被开发用于在特定发育阶段的特定组织中定位基因的增强子。在这项拟议的研究中，我们的目标是绘制在发育过程中用于内耳的增强子。了解基因在内耳中的调节方式，可能会为我们提供更多的目标，以便在我们试图恢复听力和平衡功能时进行操作。这项研究提出使用DNase I超敏反应图谱，RNA-seq和使用缺乏感觉结构域的突变小鼠来确定哪些是关键基因来完全表征增强子。这将通过以下三个具体目标来实现：目标1。在三个发育年龄绘制内耳中可接近的染色质。目标2.在缺乏感觉域的突变体中绘制DNase I超敏感区域。目标3.外植体培养中增强子活性的表征。