EFRI CEE : Engineering Technologies to Determine Causal Relationships Between Chromatin Structure and Gene Regulation

人类基因组在活细胞内的结构被精确和动态地控制，以确定不同细胞类型中每个基因的水平，以响应环境刺激和各种疾病条件。我们DNA的化学修饰和三维折叠在人类发育、疾病进展和药物反应方面发挥着比我们遗传遗传更大的作用。在绘制基因组序列和结构图方面已经取得了巨大的进展，但我们对基因组结构和功能之间的关系的了解相对较差。这项建议的目标是跨越跨学科边界，开发必要的技术，以准确预测、定量监测和确定性地编程基因组结构，以产生将催化变革性药物开发的改进的疾病模型。该团队还将开发教育项目，以激励这一新兴学科的下一代科学家。创新的新技术，包括动态分子结构记者和基于CRISPR/Cas9的表观基因组编辑，提供了一个独特的机会来监测和扰乱支配染色质结构的表观遗传状态。该团队将使用基于FRET的纳米传感器生成新的分子工具，用于监测活细胞表观遗传状态的变化，该传感器报告染色质结构和机制的结构变化，以响应基于CRISPR/Cas9的表观基因组编辑的定向扰动。这些数据将为预测表观遗传变化对染色质结构的影响以及随后的基因表达变化的分子模型提供信息。这些技术和模型将在人类肿瘤模型的背景下得到验证，以阐明染色质结构和癌症进展之间的关系。总体而言，这将使新技术和模型的开发广泛适用于疾病建模和药物筛选。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The structure of the human genome inside of a living cell is precisely and dynamically controlled to determine the level of each gene in different cell types, in response to environmental stimuli, and in various disease conditions. The chemical modification and three-dimensional folding of our DNA plays an even greater role than our inherited genetics in human development, disease progression, and drug response. There have been tremendous advances in mapping genome sequence and structure but our understanding of the relationship between genome structure and function is relatively poor. The objective of this proposal is to cross interdisciplinary boundaries to develop the necessary technologies to accurately predict, quantitatively monitor, and deterministically program genome structure to generate improved disease models that will catalyze transformative drug development. The team will also develop educational programs to inspire the next generation of scientists in this emerging discipline.Innovative new technologies, including reporters of dynamic molecular structure and CRISPR/Cas9-based epigenome editing, provide a unique opportunity to monitor and perturb epigenetic states that govern chromatin structure. The team will generate new molecular tools for monitoring changes to epigenetic states in live cells using FRET-based nanosensors that report on structural changes to chromatin structure and mechanics in response to targeted perturbations by CRISPR/Cas9-based epigenome editing. This data will inform molecular models that predict the effect of epigenetic changes on chromatin structure and consequent changes in gene expression. These technologies and models will be validated in the context of human tumor models to illuminate the relationship between chromatin structure and cancer progression. Collectively, this will enable the development of new technologies and models that are broadly useful for disease modeling and drug screening.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.