Making contact: Linking glucocorticoid receptor binding to the regulation of genes in the endogenous genomic context

建立联系：将糖皮质激素受体结合与内源基因组背景下的基因调节联系起来

• 批准号: 420008571
• 负责人: Dr. Sebastiaan H. Meijsing
• 金额: --
• 依托单位: Abteilung Bioinformatik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/420008571?language=en
• 关键词: Making; contact; Linking; glucocorticoid; receptor

Making contact: Linking glucocorticoid receptor binding to the regulation of genes in the endogenous genomic contextTranscription factors (TFs) play a pivotal role in specifying which genes are expressed in a given cell. TFs can typically bind to tens of thousands of genomic binding sites, yet they seem to regulate a much smaller number of genes. Consequently, it is mostly unclear which TF-bound regions (enhancers) are responsible for the regulation of individual genes. In work preceding this proposal, we have studied the global connection between glucocorticoid receptor (GR) binding and GR-dependent gene regulation. In addition, we studied the contribution of an individual enhancer to gene regulation using genome-editing (CRISPR/Cas9). These studies uncovered that GR binding and regulation are clearly connected, but that only a subset of binding events results in the regulation of genes. Here, we will combine genome-wide and focused studies with individual enhancers and promoters to unravel molecular mechanisms that facilitate “productive” promoter-enhancer interactions resulting in changes in gene expression. For example, we will generate and computationally analyze genome-wide data (NET-seq, Hi-ChIP, STARR-seq, etc) to identify associated candidate features (e.g. sequence motifs and 3D genome organization). In addition, the role of identified features will be studied using genome editing. One approach will be to study the effect of deleting identified sequence motifs on productive enhancer-promoter engagements. Another approach is to determine if we really understand the operating principles of productive GR binding by engineering synthetic genomic enhancers. Together, this will yield insights into the molecular mechanisms that discriminate “productive” promoter-enhancer interactions resulting in gene regulation from “non-productive” engagements.

联系：将糖皮质激素受体结合与内源性基因组环境中的基因调控联系起来转录因子（TFs）在指定哪些基因在给定细胞中表达方面起着关键作用。tf通常可以结合数以万计的基因组结合位点，但它们似乎调节的基因数量要少得多。因此，目前还不清楚哪些tf结合区域（增强子）负责调节单个基因。在此提案之前的工作中，我们研究了糖皮质激素受体（GR）结合与GR依赖基因调控之间的全局联系。此外，我们利用基因组编辑技术（CRISPR/Cas9）研究了个体增强子对基因调控的贡献。这些研究发现，GR结合和调控明显相关，但只有一部分结合事件导致基因调控。在这里，我们将结合全基因组和重点研究单个增强子和启动子，以揭示促进“生产”启动子-增强子相互作用导致基因表达变化的分子机制。例如，我们将生成和计算分析全基因组数据（NET-seq, Hi-ChIP， STARR-seq等），以确定相关的候选特征（例如序列基序和3D基因组组织）。此外，将使用基因组编辑研究已识别特征的作用。一种方法将是研究删除已识别的序列基序对产生增强子-启动子接合的影响。另一种方法是通过工程合成基因组增强子来确定我们是否真正理解了生产性GR结合的操作原理。总之，这将使我们深入了解区分导致基因调控的“生产性”启动子-增强子相互作用与“非生产性”相互作用的分子机制。