Post transcriptional control of gene expression in the lens

晶状体中基因表达的转录后控制

• 批准号: 9106633
• 负责人: Salil Lachke
• 金额: $ 39万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9106633
• 关键词: Accounting; Address; Adolescent; Alternative Splicing; Animal Model; Animals; Antibodies; Binding; Bioinformatics; Blindness; Candidate Disease Gene; Cataract; Cell Culture Techniques; Cell Cycle Regulation; Cell Differentiation process; Cells; Cellular Assay; Childhood; Complement 1q; Complex; Crystalline Lens; Cytoplasmic Granules; Data; Data Analyses; Data Set; Defect; Development; Embryonic Development; Epithelial; Epithelium; Etiology; Event; Exhibits; Eye; Family member; Fertilization; Funding; Gene Expression; Genes; Genetic; Genetic Translation; Goals; Histology; Human; Immunoprecipitation; Investigation; Knock-out; Knockout Mice; Lead; Lens Fiber; Lens development; Maintenance; Mediating; Messenger RNA; MicroRNAs; Molecular; Mutant Strains Mice; Myotonic Dystrophy; Nuclear; Online Systems; Pathogenesis; Pathway interactions; Patients; Phenotype; Post-Transcriptional Regulation; Protein Family; Proteins; Proteome; Proteomics; RNA; RNA Sequences; RNA, Messenger, Stored; RNA-Binding Proteins; Regulation; Research; Scanning Electron Microscopy; Scientist; Staging; System; Tertiary Protein Structure; Testing; Tissues; Transcript; Translational Repression; Translations; base; early onset; fiber cell; gene discovery; gene function; insight; interactive tool; kinase inhibitor; lens; lens transparency; mRNA Stability; mutant; novel; protein function; public health relevance; research study; tool; transcription termination; transcriptome; transcriptome sequencing; user-friendly

 DESCRIPTION (provided by applicant): Cataract, the opacification of the ocular lens, is the leading cause of blindness worldwide. The etiology of most cases of sporadic pediatric cataract is still unknown, and genetic anomalies are estimated to account for 25-50% of these cases. We have developed a novel bioinformatics approach, iSyTE (integrated Systems Tool for Eye gene discovery, http://bioinformatics.udel.edu/Research/iSyTE) to predict genes that are associated with cataract. In the recent past, our finding that deficiency of the RNA granule (RG) component TDRD7 causes juvenile cataracts in human and animal models have indicated the significance of post-transcriptional regulatory mechanisms in the development and maintenance of lens transparency. We have now used iSyTE to identify a new conserved RNA binding protein and RG component Celf1 whose germline or lens-specific deletion in mouse mutants causes early onset cataracts. In this proposal, we will test the overarching hypothesis that Celf1 mediates post-transcriptional control of gene expression to regulate lens development. Specifically, we will address the following goals. (Aim 1) Characterize the pathogenesis of lens defects in Celf1 mouse mutants and gain insights into the molecular underpinning of lens defects in Celf1 mutants by analysis of the lens transcriptome and proteome. (Aim 2) Elucidate the direct RNA targets of Celf1 by RNA-immunoprecipitation followed by RNA-Sequencing (RNA- Seq). Further, test the mechanism of Celf1-mediated regulation for high-priority candidate genes that function in the lens, and are selected based on stringent filtering criteria. Specifically, we will investigate the molecular mechanism of Celf1 function in: 1) translational control of the cell cycl kinase inhibitor p27, 2) control of Prox1 expression in lens epithelium, and 3) control of mRNA stability for the fiber cell differentiation factor essential for nuclear degradation, Dnase2b. (Ai 3) Finally, integrate and analyze these data on Celf1 within the context of existing lens data to derive Celf-regulatory networks in the lens. These investigations aimed at elucidating the mechanism of Celf1 function will advance the understanding of post-transcriptional control of gene expression in the lens and lead to identification of new targets associated with cataracts.

 描述（由申请人提供）：白内障，即晶状体混浊，是全世界失明的主要原因。大多数散发性儿童白内障病例的病因仍不清楚，估计遗传异常占这些病例的 25-50%。我们开发了一种新颖的生物信息学方法 iSyTE（眼基因发现集成系统工具，http://bioinformatics.udel.edu/Research/iSyTE）来预测与白内障相关的基因。最近，我们发现人类和动物模型中RNA颗粒（RG）成分TDRD7的缺乏会导致青少年白内障，这表明转录后调节机制在晶状体透明度的发育和维持中的重要性。我们现在使用 iSyTE 鉴定了一种新的保守 RNA 结合蛋白和 RG 成分 Celf1，其种系或晶状体特异性缺失在小鼠突变体中会导致早发性白内障。在本提案中，我们将测试 Celf1 介导基因表达转录后控制以调节晶状体发育的总体假设。具体来说，我们将实现以下目标。 （目标 1）表征 Celf1 小鼠突变体晶状体缺陷的发病机制，并通过分析晶状体转录组和蛋白质组深入了解 Celf1 突变体晶状体缺陷的分子基础。 （目标 2）通过 RNA 免疫沉淀和 RNA 测序 (RNA-Seq) 阐明 Celf1 的直接 RNA 靶标。此外，测试 Celf1 介导的对晶状体中起作用的高优先级候选基因的调节机制，并根据严格的过滤标准进行选择。具体来说，我们将 研究 Celf1 功能的分子机制：1) 细胞周期激酶抑制剂 p27 的翻译控制，2) 晶状体上皮中 Prox1 表达的控制，以及 3) 核降解所必需的纤维细胞分化因子 Dnase2b 的 mRNA 稳定性的控制。 (Ai 3) 最后，在现有晶状体数据的背景下整合和分析 Celf1 上的这些数据，以得出晶状体中的 Celf 调节网络。这些旨在阐明 Celf1 功能机制的研究将增进对晶状体基因表达转录后控制的理解，并导致与白内障相关的新靶点的识别。