Genome-wide location analysis of neuronal genes

神经元基因的全基因组定位分析

• 批准号: 6968031
• 负责人: MICHAEL G ROSENFELD
• 金额: $ 30.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6968031
• 关键词: biological signal transduction; calmodulin dependent protein kinase; chromatin immunoprecipitation; cofactor; developmental neurobiology; enzyme linked immunosorbent assay; gene induction /repression; genetically modified animals; genome; genotype; laboratory mouse; neurogenetics; nucleic acid hybridization; small interfering RNA; technology /technique development; transcription factor

DESCRIPTION (provided by applicant): The precisely-regulated pattern of recruitment of DNA binding transcription factors and associated coactivators and corepressors underlies the critical events that regulate neurodevelopment and disease. We propose to validate 3 distinct technologies that will permit genome-wide promoter location analysis for these regulatory factors-ChlP-on-chip, ChlP-DASL-chip, and ChlP-RDA-for applications to test specific hypotheses in neurodevelopment and disease. Genome-wide promoter assays will be designed, constructed and validated and we will test these technologies by determining the pattern of cofactor binding in the genome. New siRNA approaches to profiling will be applied to validate occupied promoters as functional targets. We will focus on a method that provides the optimal sensitivity and applicability to gene tiling, and will generate a genome-wide murine promoter array and will tile a series of genomic intervals that will permit investigation of specific genomic loci to uncover epigenetic strategies used in signaldependent changes in gene activation/repression programs development. These include study of the roles of CaMKII delta and SMRT in early programs of neurodevelopment. This approach will permit testing of several hypotheses and will be used to identify the promoter-specific usage of key corepressors and coactivators and location of epigenetic marks. We believe that with validation of these approaches, technologies developed here will be of widespread utility in neuroscience, particularly in studies of development and neurodegenerative disease.

描述（由申请人提供）：DNA 结合转录因子以及相关的共激活因子和辅抑制因子的精确调控模式是调节神经发育和疾病的关键事件的基础。我们建议验证 3 种不同的技术，这些技术将允许对这些调节因子（ChlP-on-chip、ChlP-DASL-chip 和 ChlP-RDA）进行全基因组启动子定位分析，用于测试神经发育和疾病中的特定假设。将设计、构建和验证全基因组启动子测定，我们将通过确定基因组中辅因子结合的模式来测试这些技术。新的 siRNA 分析方法将用于验证占据的启动子作为功能靶点。我们将重点关注一种为基因平铺提供最佳灵敏度和适用性的方法，并将生成全基因组鼠启动子阵列，并将平铺一系列基因组间隔，这将允许研究特定基因组位点，以揭示基因激活/抑制程序开发中信号依赖性变化中使用的表观遗传策略。其中包括 CaMKII delta 和 SMRT 在神经发育早期程序中的作用的研究。这种方法将允许测试几个假设，并将用于识别关键辅抑制子和共激活子的启动子特异性使用以及表观遗传标记的位置。我们相信，通过验证这些方法，这里开发的技术将在神经科学中广泛应用，特别是在发育和神经退行性疾病的研究中。