A Platform Technology for High-Throughput Screening of Gene Regulatory Elements

项目摘要/摘要 这项提案的目标是开发一个新的高通量平台来理解基因调控 要素，以确定针对常见疾病的新药靶点。人类基因组编码 大约50,000个基因。了解这些基因是如何被调控的，以及这与 长期以来，复杂的细胞表型一直是我们团队的主要关注点。人类基因组的后续项目 项目，如NIH资助的DNA元素百科全书(ENCODE)和路线图表观基因组学 项目已经确定了许多人类细胞在人类基因组中可能存在的数百万个调控元件 类型和组织。重要的是，基因组广泛关联(GWA)研究已经强烈表明，非 编码调控元件决定了导致大多数复杂疾病的基因表达模式 包括癌症、心血管疾病、糖尿病和神经疾病。然而，这些功能的功能 调控元件及其与这些疾病表型的关系在很大程度上是未知的。另外， 干扰细胞过程的常规筛选技术，如小分子和RNA 干扰，不能直接针对基因组调控元件。要解决这一关键限制，并 阐明了这些细胞表型的基本基因组基础，我们最近开发了表观基因组- 直接和精确地激活和抑制其基因组调控元件的编辑技术 自然染色体位置。最近，我们开发了一种新的和健壮的方法来使用这些 高通量鉴定和量化基因调控元件活性的工具。在这里，我们建议 将这些方法应用于发现和验证与癌症和 心血管疾病作为了解遗传基础的新平台技术的例证 一种复杂的疾病。这项技术将对转化遗传学和基因组学的现代进步至关重要 进入新的药物靶点、诊断和个性化药物以迎合每个患者的基因组。

Project Summary/Abstract The goal of this proposal is to develop a novel high-throughput platform for understanding gene regulatory elements in order to identify new drug targets for common diseases. The human genome encodes approximately 50,000 genes. Understanding how those genes are regulated and how this correlates to complex cell phenotypes has long been a major focus of our team. Follow-up projects to the Human Genome Project, such as the NIH-funded Encyclopedia of DNA Elements (ENCODE) and the Roadmap Epigenomics Project, have identified millions of putative regulatory elements across the human genome for many human cell types and tissues. Importantly, genome wide association (GWA) studies have strongly indicated that non- coding regulatory elements determine the gene expression patterns responsible for most complex diseases including cancer, cardiovascular disease, diabetes, and neurological disorders. However, the function of these regulatory elements and their relationships to these disease phenotype are largely unknown. Additionally, conventional screening technologies for perturbing cellular processes, such as small molecules and RNA interference, cannot directly target genomic regulatory elements. To address this critical limitation and illuminate the fundamental genomic basis of these cell phenotypes, we have recently developed epigenome- editing technologies for directly and precisely activating and repressing genomic regulatory elements in their natural chromosomal location. More recently, we have developed a novel and robust method for using these tools for high-throughput identification and quantification of gene regulatory element activity. Here, we propose to apply these methods to the discovery and validation of regulatory elements associated with cancer and cardiovascular disease as demonstration of this novel platform technology for understanding the genetic basis of complex disease. This technology will be critical to translating modern advances in genetics and genomics into new drug targets, diagnostics, and personalized medicine catered to each patient genome.