Optogenetic Engineering of 3D Chromatin Architecture

项目摘要/摘要 染色质三维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级方面取得了快速的进展， 包括增强子-启动子环、染色质接触域、拓扑域和染色质A/B组分。 分部。重要的是，3D基因组结构的改变在人类疾病中扮演着重要的角色 比如发育障碍和癌症。然而，当前的一个主要挑战是确定生物- 计算这些3D染色质结构的每一层的重要性，以揭示它们在调节- ING基因表达和/或其他核活动，并潜在地操纵这些结构以进行分布。 放松干预。我们组建了一支强大的调查团队来开发一种创新的新工具集来帮助 解决这一挑战，它利用了光可控的蛋白质二聚体和先进的Crispr/Cas9 GE- 诺姆编辑技术。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来创建从头开始的增强子-启动子环和拓扑相关结构域，这可能会迅速 激活和停用。该项目的成功实施将提供一种非常必要的战略，以 在体外和体内研究染色质基本上在任何特定的感兴趣区域的环。这个工具集速度很快， 高效和非侵入性，这不仅将为揭示染色质的深刻新见解铺平道路 基因转录控制中的环和结构域，但也揭示了一个纠正疾病的新概念- 染色质结构的相关畸形形成。

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.