Elucidating factors that modulate mammalian DNA repair to improve genome editing

项目总结/摘要 基因组编辑是治疗常见和罕见遗传疾病的令人兴奋的途径， CRISPR/Cas技术加速了这种治疗选择的发展。这些疗法依赖于 细胞DNA修复机制来安装它们的编辑，因此改变了DNA修复途径之间的平衡 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 在细胞类型之间或甚至在同一细胞中的目标之间的编辑效率。因此，我的长期目标是 该实验室的目标是研究改变基因组编辑功效的因素，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，组蛋白修饰与偏见有关 特定的DNA修复途径。然而，考虑到它们在多个过程中的作用， 个体组蛋白修饰一直是一个挑战。我将剖析这些影响，以确定组蛋白的作用， 通过创新一个平台，招募组蛋白修饰剂， 数以千计的断裂点这项工作将确定组蛋白修饰是否影响DNA修复 并揭示预测这些效应的目标部位的特性。基因组编辑要达到治疗效果 需要潜在的、精确的DNA修复控制。我会从人类蛋白质组中找出能改变 修复双链断裂为了做到这一点，我将采用我开发的肽筛选平台来研究 DNA修复这些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 修复.这些项目的完成将大大推进我们对哺乳动物DNA修复的理解 并有可能改善基因组编辑疗法。

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.