High throughput interrogation of non-coding variants and 3D genome organization

非编码变异和 3D 基因组组织的高通量询问

基本信息

批准号：
10669105
负责人：
Feng Yue
金额：
$ 40.8万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2027-07-31
项目状态：
未结题

项目摘要

PROJECT SUMMARY / ABSTRACT Recent large-scale efforts such as the ENCODE and Epigenome Roadmap projects have predicted millions of potential non-coding regulatory elements in the human genome. However, it is not clear how many of them are truly functional, as the predictions were mainly based on high-throughput genomics assays such as DNase- Seq or ChIP-Seq for histone modifications and transcription factors. Most of the current studies indirectly measure enhancer activity through transgenic reporter assays and therefore do not provide insights into the native chromatin state. Fortunately, recent advances in high-throughput screening methods based on CRISPR/Cas9 genome editing technology make such functional characterization possible. Another important layer of gene regulation is the 3D genome organization, which can link the distal enhancers with their target genes. High-throughput methods based on Chromatin Conformation Capture (3C) have emerged (such as Hi- C, ChIA-PET, HiChIP, and Capture Hi-C) and present an unprecedented opportunity to study higher-order chromatin structure genome-wide. Despite the recent advances, the complex relationship between chromatin interactions and gene regulation has just begun to unravel. CTCF and cohesin complex have been shown to be critical in the formation of chromatin loops and topologically associating domains (TADs), but we know little of whether there are other important regulators are important for such chromatin interactions. Given the aforementioned challenges and my unique multi-disciplinary training, my long-term goal is to use a combination of high throughput genomic experiments, computational modeling, and functional assays to address the following two fundamental questions: 1) Identify functional enhancers through high-throughput assays and study how genetic variants can affect their functions; 2) Identify novel regulators for the formation of 3D genome organization features. The proposed work will deepen our understanding on how genetic variants contribute to gene regulation, 3D genome organization, and molecular mechanisms underlying human diseases.

项目摘要 /摘要最近的大规模努力（例如编码和表观基因组路线图项目）预测了数百万人类基因组中潜在的非编码调节元件。但是，尚不清楚其中有多少真正的功能，因为这些预测主要基于高通量基因组学测定，例如DNase- 用于组蛋白修饰和转录因子的SEQ或CHIP-SEQ。当前的大多数研究间接研究通过转基因记者测定法测量增强子活性，因此没有提供有关天然染色质状态。幸运的是，基于高通量筛选方法的最新进展 CRISPR/CAS9基因组编辑技术使这种功能表征成为可能。另一个重要基因调节层是3D基因组组织，它可以将远端增强子与目标联系起来基因。已经出现了基于染色质构象捕获（3C）的高通量方法（例如HI- c，chia-pet，hichip和Capture hi-c），并提供了一个前所未有的机会来研究高阶染色质结构全基因组。尽管最近取得了进步，但染色质之间的复杂关系相互作用和基因调节刚刚开始解散。 CTCF和粘蛋白复合物已显示为在形成染色质环和拓扑结构域（TADS）中至关重要，但我们知道很少是否还有其他重要调节剂对于这种染色质相互作用很重要。鉴于上述挑战和我独特的多学科培训，我的长期目标是使用高通量基因组实验，计算建模和功能测定的组合解决以下两个基本问题：1）通过高通量确定功能增强器测定和研究遗传变异如何影响其功能； 2）确定形成的新型调节剂 3D基因组组织功能。拟议的工作将加深我们对遗传的理解变体有助于人类基因调节，3D基因组组织和分子机制疾病。