In vivo multiplexed silencing of cis-elements in the brain

项目摘要 阿尔茨海默氏病（AD）是美国的一种主要的个人和公共健康危机，其发病率是 增加。代谢和炎症性疾病通过驱动基因调节和 表情变化。事实上，衰老过程中的表观遗传学变化可能揭示出重要的治疗靶点， 预防AD。我们工作的长期目标是揭示核心顺式元件和基因调控网络 （GRN）控制AD风险。一个挑战是GRNs涉及多个顺式调控元件和基因 在基因组中。目前，缺乏在体内研究顺式元件组合的功能的方法 使用小鼠模型，这是一个障碍，阻止我们识别可以预防或解决AD的GRNs 病理犹他州大学的格雷格和格茨实验室合作研究的目标是 是在小鼠中开发一种方法来研究体内顺式元件组合的功能。的 该项目将为GRNs的功能研究创建一个平台技术， 保守的GRN可以预防或解决AD病理。该方法可以应用于许多 生物问题。在最近的Cell Systems论文中，我们（Gertz实验室）设计了一种基于CRISPR的技术， 能够使多个增强子同时表观遗传失活。这种方法被称为增强器- 干扰（增强子-I），并且可以同时维持多达50个基因座的稳定沉默。增强剂-I是 用于体外研究的增强子-I系统和用于小鼠模型中的体内表观基因组编辑的增强子-I系统不 仍然存在因此，我们正在与Gregg实验室合作，使增强子-I适应体内工作。格雷格实验室 在研究小鼠表观遗传学和基因调控方面拥有丰富的经验， 揭示了保守的顺式元件和GRNs的图谱，这些元件和GRNs是控制相关过程的候选者。 包括肥胖、炎症和神经变性。在这里，我们将合作创建一个鼠标 用于多重体内表观基因组编辑的增强子-I系统的模型。为了证明原理，我们的目标是 顺式元件控制Tau、β-淀粉样前体蛋白（App）、脂肪质量和肥胖（FTO）基因座和 其他重要的AD风险位点。增强子-I将使研究人员能够研究顺式- 元件和定义控制体内AD病理学的功能性GRNs。我们的技术将帮助开辟新的领域 学习。

PROJECT SUMMARY Alzheimer’s disease (AD) is a major personal and public health crisis in the United States and the incidence is increasing. Metabolic and inflammatory disorders contribute to AD risks by driving gene regulation and expression changes. Indeed, epigenetic changes during aging could reveal important therapeutic targets for preventing AD. The long-term goal of our work is to uncover the core cis-elements and gene regulatory networks (GRNs) that control AD risks. A challenge is that GRNs involve multiple cis-regulatory elements and genes across the genome. Currently, methods are lacking to functionally study combinations of cis-elements in vivo using mouse models, which is a barrier blocking us from identifying GRNs that can prevent or resolve AD pathology. The goal of this collaborative study between the Gregg and Gertz labs at the University of Utah is to develop an approach in mice to study the functions of combinations of cis-elements in vivo. The project will create a platform technology for functional studies of GRNs, facilitating the identification of conserved GRNs that can prevent or resolve AD pathology. The approach can be applied to many biological problems. In a recent Cell Systems paper, we (Gertz lab) devised a CRISPR- based technique that enables simultaneous epigenetic deactivation of multiple enhancers. The method is called Enhancer- interference (Enhancer-I) and can maintain stable silencing of up to 50 loci simultaneously. Enhancer-I was developed for in vitro studies and an Enhancer-I system for in vivo epigenome editing in mouse models does not yet exist. Therefore, we are collaborating with the Gregg lab to adapt Enhancer-I for in vivo work. The Gregg lab has extensive experience studying epigenetics and gene regulation in mice and recently published studies uncovering an atlas of conserved cis-elements and GRNs that are candidates for controlling processes involved in AD, including obesity, inflammation and neurodegeneration. Here, we will collaborate to create a mouse model of the Enhancer-I system for multiplexed in vivo epigenome editing. For proof-of-principle, we target cis-elements controlling Tau, beta-amyloid precursor protein (App), the Fat Mass & Obesity (FTO) locus and other important AD risk loci. Enhancer-I will empower researchers to study the functions of combinations of cis- elements and define functional GRNs controlling AD pathology in vivo. Our technology will help open new areas of study.