Elucidating factors that modulate mammalian DNA repair to improve genome editing

阐明调节哺乳动物 DNA 修复的因素以改善基因组编辑

• 批准号: 10712484
• 负责人: Gaelen T Hess
• 金额: $ 38.88万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712484
• 关键词: Acceleration; Address; CRISPR/Cas technology; Cell physiology; Cells; DNA Repair; DNA Repair Pathway; Dependence; Double Strand Break Repair; Equilibrium; Genetic Diseases; Goals; Health; Histones; Human; Individual; Laboratories; Methods; Mission; Outcome; Pattern; Peptides; Process; Property; Proteome; Public Health; Research; Role; Site; Therapeutic; Transcriptional Regulation; United States National Institutes of Health; Work; cell type; genome editing; histone modification; improved; innovation; protein protein interaction; rare genetic disorder; recruit; screening; therapeutic development; therapeutic genome editing

Project Summary/Abstract Genome editing is an exciting avenue for treating common and rare genetic diseases, and the advent of CRISPR/Cas technologies has accelerated the development of this therapeutic option. These therapies rely on cellular DNA repair machinery to install their edits, so changes in the balance between DNA repair pathways result in different editing patterns. This dependency on endogenous DNA repair has led to considerable variability in editing efficiency between cell types or even among targets in the same cell. Thus, a long-term goal of my laboratory is to investigate factors that modify the efficacy of genome editing and to develop strategies that address these shortcomings. Histone modifications play a role in many cellular processes, including transcriptional regulation and DNA repair. Recent evidence found that histone modifications correlate with biases for specific DNA repair pathways. However, given their role in multiple processes, interpreting the effects of individual histone modifications has been challenging. I will dissect these effects to define the role of histone modifications in the repair of double-strand breaks by innovating a platform that recruits histone modifiers to thousands of break sites in parallel. This work will determine whether histone modifications influence DNA repair and uncover properties of the target site that predict these effects. For genome editing to reach its therapeutic potential, precise control of DNA repair is required. I will identify peptides from the human proteome that alter the repair of double-strand breaks. To do this, I will adapt a peptide screening platform I developed to investigate DNA repair. The peptides will reveal critical protein-protein interactions and other methods for altering DNA repair. The completion of these projects will significantly advance our understanding of mammalian DNA repair and has the potential to improve genome editing therapies.

项目摘要/摘要 基因组编辑是治疗常见和罕见遗传病的一种令人兴奋的途径，而 CRISPR/CAS技术加速了这一治疗方案的发展。这些疗法依赖于 细胞DNA修复机制安装它们的编辑，使DNA修复途径之间的平衡发生变化 导致不同的编辑模式。这种对内源性DNA修复的依赖导致了相当大的变异性 在单元类型之间甚至同一单元中的目标之间的编辑效率方面。因此，我的一个长期目标是 实验室的任务是调查改变基因组编辑效果的因素，并制定策略 解决这些缺点。组蛋白修饰在许多细胞过程中发挥作用，包括 转录调控和DNA修复。最近的证据发现，组蛋白修饰与偏向有关 用于特定的DNA修复途径。然而，考虑到它们在多个过程中的作用，解释 单独的组蛋白修饰一直是具有挑战性的。我将剖析这些影响，以确定组蛋白的作用 通过创新一个招募组蛋白修饰剂的平台来修改双链断裂的修复 数以千计的休息点并行。这项工作将确定组蛋白修饰是否会影响DNA修复 并揭示预测这些影响的目标站点的属性。使基因组编辑达到其治疗效果 需要对DNA修复进行潜在的、精确的控制。我将从人类蛋白质组中鉴定出可以改变 双链断裂的修复。为此，我将采用我开发的一个肽筛选平台来研究 DNA修复。这些多肽将揭示关键的蛋白质-蛋白质相互作用和其他改变DNA的方法 修理。这些项目的完成将极大地促进我们对哺乳动物DNA修复的理解 并有可能改进基因组编辑疗法。