Deciphering the contribution of enhancer transcription to enhancer function

破译增强子转录对增强子功能的贡献

• 批准号: 10316087
• 负责人: Carlos Guzman
• 金额: $ 3.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10316087
• 关键词: Deciphering; contribution; enhancer; transcription; function

PROJECT SUMMARY/ABSTRACT The applicant is requesting two years of support from a Ruth L. Kirschstein National Research Service Award for Individual Predoctoral Fellowship to Promote Diversity in Health-Related Research (F31-Diversity) to examine the relationship between enhancer transcription and enhancer function. Enhancers have been shown to initiate short, non-coding transcripts in quantities rivaling promoters, yet their role on enhancer function remains controversial. My preliminary data suggests that enhancers and promoters synchronize their activity to form co- regulated domains. This result is surprising given that previous studies using total RNA measurements have indicated that enhancer eRNAs ramp up before their associated mRNAs, suggesting that enhancers get activated first and that this activation then gets transmitted to the associated promoters. Co-regulated domains explain several of the mechanisms that the textbook model of enhancer function fails to. These include that enhancer sequences are indistinguishable from promoters, that they are bound and initiate transcription by RNA polymerase II, that enhancers and promoters form hubs in the nucleus, that enhancers act in an additive or synergistic manner, and the redundancies observed when knocking out single enhancers in a multi-enhancer locus. The goal of this research is to test the hypothesis that enhancers may function by serving as simultaneously activated and mutually reinforcing transcription initiation sites within CRDs. This hypothesis implies that enhancer transcription would be essential for enhancer function. I will test this hypothesis in three specific aims: (1) determine the generality of enhancer-promoter co-regulatory domains across signaling responses, to uncover whether the formation of co-regulated domains are a general regulatory mechanism underlying signal response (2) determine the relationship between enhancer transcription and enhancer function outside of their native genomic environment, to examine the relationship between enhancer transcription and enhancer function outside of their native genomic context, and (3) determine the impact of silencing enhancer transcription on gene activation in vivo, to determine the influence that shutting down enhancer transcription ahs on enhancer function in its native genomic context. This training will allow me to (1) develop skills in the areas of research design, analysis and interpretation using next-generation sequencing and molecular biology techniques; (2) learn the fundamentals of molecular, chromatin, and enhancer biology; (3) successfully defend a dissertation; and (4) obtain a competitive post-doctoral fellowship with the long-term goal to become a successful, independently-funded scientist at a research-intensive university.

项目总结/摘要 申请人要求露丝·L·史密斯提供两年的支持。基尔施斯坦国家研究服务奖 个人博士前奖学金，以促进健康相关研究的多样性（F31-多样性），以审查 增强子转录和增强子功能之间的关系。增强剂已被证明可以启动 短的，非编码转录本的数量与启动子相媲美，但它们对增强子功能的作用仍然存在 争议我的初步数据表明，增强子和启动子同步它们的活动，形成共同的， 规范的领域。这一结果令人惊讶，因为以前使用总RNA测量的研究 表明增强子eRNAs在其相关mRNA之前上升，表明增强子 首先激活，然后将这种激活传递给相关的启动子。共调控域 解释增强子功能的教科书模型未能解释的几种机制。其中包括， 增强子序列与启动子是不可区分的，它们被RNA结合并启动转录。 聚合酶II，增强子和启动子在细胞核中形成枢纽，增强子以添加剂或 协同方式，以及当敲除多增强子中的单个增强子时观察到的冗余 基因座这项研究的目的是测试这一假设，即增强子可能通过充当 同时激活并相互增强CRD内的转录起始位点。这一假设 这意味着增强子转录对于增强子功能是必需的。我将在三个方面来检验这个假设 具体目的：（1）确定增强子-启动子共调控结构域在信号转导中的一般性 反应，以揭示共调节域的形成是否是一种普遍的调节机制 潜在的信号响应（2）决定增强子转录和增强子功能之间的关系 在它们的天然基因组环境之外，检查增强子转录和 增强子在其天然基因组背景之外的功能，以及（3）确定沉默增强子的影响。 转录对体内基因激活的影响，以确定关闭增强子转录AHS 增强子在其天然基因组环境中的功能。这项培训将使我能够（1）在以下领域发展技能 使用下一代测序和分子生物学进行研究设计、分析和解释 技术;（2）学习分子，染色质和增强子生物学的基础知识;（3）成功地捍卫 论文;以及（4）获得具有竞争力的博士后奖学金，长期目标是成为 在一所研究密集型大学获得独立资助的成功科学家。