In vivo multiplexed silencing of cis-elements in the brain

项目概要 阿尔茨海默病（AD）是美国的一个重大个人和公共健康危机，发病率 增加。代谢和炎症性疾病通过驱动基因调控和 表情发生变化。事实上，衰老过程中的表观遗传变化可能揭示重要的治疗靶点 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。挑战在于 GRN 涉及多个顺式调控元件和基因 跨越基因组。目前，缺乏对体内顺式元件组合进行功能研究的方法 使用小鼠模型，这是我们识别可以预防或解决 AD 的 GRN 的障碍 病理学。犹他大学 Gregg 和 Gertz 实验室合作研究的目标 旨在开发一种在小鼠体内研究顺式元件组合功能的方法。的 该项目将创建一个用于GRN功能研究的平台技术，促进GRN的识别 保守的 GRN 可以预防或解决 AD 病理。该方法可以应用于许多 生物学问题。在最近的一篇细胞系统论文中，我们（格茨实验室）设计了一种基于 CRISPR 的技术 使多个增强子同时表观遗传失活。该方法称为Enhancer- 干扰（增强子-I），并且可以同时维持多达 50 个位点的稳定沉默。增强剂-我是 为体外研究开发的和用于小鼠模型体内表观基因组编辑的Enhancer-I系统不 但仍然存在。因此，我们正在与 Gregg 实验室合作，将Enhancer-I 应用于体内工作。格雷格实验室 拥有研究小鼠表观遗传学和基因调控的丰富经验以及最近发表的研究 揭示保守顺式元素和 GRN 的图集，它们是控制相关过程的候选者 AD，包括肥胖、炎症和神经退行性疾病。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Enhancer-I系统模型。为了进行原理验证，我们的目标是 顺式元件控制 Tau、β-淀粉样前体蛋白 (App)、脂肪量和肥胖 (FTO) 位点和 其他重要的 AD 风险位点。增强剂-I 将使研究人员能够研究顺式组合的功能 元素并定义控制体内 AD 病理的功能性 GRN。我们的技术将帮助开辟新领域 的学习。

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.