Optogenetic Engineering of 3D Chromatin Architecture

项目总结/摘要 三维染色质结构的组织原理仍然是生物学中的一个主要谜团。 在过去的十年中，在表征染色质结构的层次结构方面取得了快速进展， 包括增强子-启动子环、染色质接触结构域、拓扑结构域和染色质A/B com- - 是的重要的是，3D基因组结构的改变在人类疾病中起着重要作用 例如发育障碍和癌症。然而，目前的一个主要挑战是确定生物学- 这些3D染色质结构的每一层的重要性，以揭示它们在调节中的致病作用， 研究基因表达和/或其他核活动，并潜在地操纵这些结构以用于研究。 方便干预。我们组建了一支强大的调查团队，开发了一套创新的新工具， 解决这一挑战，它利用光可控蛋白二聚体和先进的Crispr/Cas9 ge- nome编辑技术。我们在这个项目中的目标是开发我们提到的这个工具集的原型 作为PaCIR来创建从头增强子-启动子环和拓扑相关结构域， 激活和停用。该项目的成功实施将提供一个高度要求的战略， 在体外和体内研究基本上任何感兴趣的特定区域的染色质环。这个工具集是快速的， 高效和非侵入性，这不仅为揭示染色质的深刻新见解铺平了道路， 环和结构域在基因转录控制，而且还揭示了一个新的概念，以纠正疾病- 相关的染色质结构畸形。

PROJECT SUMMARY / ABSTRACT The organizing principle of chromatin architecture in three dimensions (3D) remains a major mystery in biology. A rapid progress has been made in the last decade to characterize the hierarchies of chromatin architecture, including enhancer-promoter loops, chromatin contact domains, topological domains and chromatin A/B com- partments. Importantly, the alteration of the 3D genome architecture plays an important role in human disease such as developmental disorders and cancer. However, a major current challenge is to determine the biologi- cal importance of each layer of these 3D chromatin architectures, to uncover their causative roles in modulat- ing gene expression and/or other nuclear activities, and to potentially manipulate these architectures for dis- ease intervention. We assembled a strong team of investigators to develop an innovative new toolset to help solve this challenge, which takes advantage of light controllable protein dimers and advanced Crispr/Cas9 ge- nome editing technology. Our goal in this project is to develop the prototype of this toolset that we referred to as PaCIR to create de novo enhancer-promoter loops and topological associated domain that could be rapidly activated and deactivated. The successful execution of this project will provide a highly demanded strategy to study chromatin loops at essentially any specific regions of interests in vitro and in vivo. This toolset is rapid, efficient and non-invasive, which will not only pave way for uncovering profound new insights into chromatin loops and domains in gene transcription control, but also shed lights on a novel concept to rectify disease- associated mal-formation of chromatin architectures.