Programmable DNA loops between PU.1 gene regulatory elements

PU.1 基因调控元件之间的可编程 DNA 环

• 批准号: 428912142
• 负责人: Professor Dr. Frank Rosenbauer
• 金额: --
• 依托单位: Institut für Molekulare Tumorbiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/428912142?language=en
• 关键词: Programmable; DNA; loops; between; PU.1

Disruption of spatial chromosomal architecture is an emerging mechanism in human disease, thus presenting a yet unexplored potential therapeutic option. However, the spatial chromosomal requirements of most genes are yet unknown, especially of those involved in diseases. The ETS-domain transcription factor PU.1 is essential for the production of myeloid and lymphoid progenitor cells and disturbed PU.1 expression can lead to leukemia. Previous work by us and others revealed that PU.1 expression is under tight control of a multilayered interplay of lineage-specific transcription factors with distal gene regulatory elements. Furthermore, we have recently published high-resolution chromosome conformation maps revealing the three-dimensional organization of the PU.1 locus in both healthy and leukemic cells. Our work linked PU.1 autoregulation with specific long-range intrachromosomal contacts, and correlated dynamic chromatin looping with PU.1 expression. With the present proposal, we will engineer reversible chromatin loops between different PU.1 regulatory elements by a new CRISPR-dCas9-based method, and thus will functionally access involvement of the three-dimensional chromosomal architecture in driving lineage-specific and disease-associated PU.1 expression levels. Our work will provide causal information on how the spatial chromatin landscape orchestrates myeloid differentiation and malignant transformation.

空间染色体结构的破坏是人类疾病中的一种新兴机制，因此提出了一种尚未探索的潜在治疗选择。然而，大多数基因的空间染色体要求是未知的，特别是那些涉及疾病。ETS结构域转录因子PU.1对于髓样和淋巴样祖细胞的产生是必需的，并且PU.1表达紊乱可导致白血病。我们和其他人以前的工作表明，PU.1的表达是在一个多层的相互作用的谱系特异性转录因子与远端基因调控元件的严格控制。此外，我们最近发表了高分辨率的染色体构象图，揭示了健康和白血病细胞中PU.1位点的三维组织。我们的工作将PU.1自动调节与特定的长距离染色体内接触联系起来，并将动态染色质循环与PU.1表达联系起来。通过本提案，我们将通过一种新的基于CRISPR-dCas 9的方法在不同的PU.1调控元件之间设计可逆的染色质环，从而在功能上访问三维染色体结构在驱动谱系特异性和疾病相关的PU.1表达水平中的参与。我们的工作将提供空间染色质景观如何编排骨髓分化和恶性转化的因果信息。