MutSensor System: A Set of Highly Sensitive Mutation Reporters to Dissect Genome Stability in Health and Disease

MutSensor 系统：一组高度灵敏的突变报告基因，用于剖析健康和疾病中基因组的稳定性

• 批准号: 10737167
• 负责人: Jef D BOEKE
• 金额: $ 75.84万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10737167
• 关键词: Binding; Biological Assay; Carcinogens; Cell Line; Cells; Chromosome Mapping; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Communities; DNA Damage; DNA Repair; DNA Sequence Alteration; DNA cassette; Data; Disease; Event; Exposure to; Family; Frequencies; Genes; Genetic; Genetic Determinism; Genetic Screening; Genetic study; Genome; Genome Stability; Germ-Line Mutation; Goals; Guide RNA; HCT116 Cells; Health; Human; Human body; Individual; Induced Mutation; Knowledge; Laboratories; Lead; Left; Libraries; Location; Malignant Neoplasms; Mammalian Cell; Methodology; Methods; Mismatch Repair; Mutagenesis; Mutagens; Mutate; Mutation; Mutation Detection; Open Reading Frames; Pathway interactions; Pharmaceutical Preparations; Point Mutation; Process; Proteins; Reporter; Repression; Resistance; Shapes; Signal Transduction; Single Nucleotide Polymorphism; Site; Sorting; System; Testing; Time; Tissues; Variant; Writing; Yeasts; carcinogenesis; cell type; design; experimental study; follow-up; gain of function; genetic approach; genome integrity; genome-wide; human disease; human tissue; innovation; loss of function; loss of function mutation; model organism; mutant; novel; overexpression; promoter; prototype; screening; sensor; tool; variant detection

Summary Our cells are constantly exposed to mutagens that cause DNA damage, which if left unrepaired, cause mutations. At the cellular level, the progressive accumulation of mutations in somatic tissues drives carcinogenesis and other diseases. Despite significant advances in our understanding of mutational processes, the answer to many fundamental questions is still a mystery. What are the genetic causes of mutations in human tissues? How do different cell types in the human body control DNA damage and repair? Despite our large body of knowledge in the pathways that control DNA damage and repair in model organisms, we lack a deep understanding of this process in human tissues. This would be critical to understand how mutations lead to human diseases including carcinogenesis. Here we will use a highly innovative approach to build MutSensor, a set of mutation reporters to estimate DNA mutation frequency in mammalian cells with a sensitivity >50-fold higher than that of existing methods. The ability to build and precision deliver these MutSensors is enabled by “Genetic Writing and Delivery” system developed in the Boeke lab. This novel method will allow large-scale functional genetic screening at an unprecedented scale. Building on our preliminary data, we aim at comprehensively identify all genes that regulate mutation frequency in different human cell types. To this purpose, we will utilize loss- and gain-of-function screening libraries to determine the effect of genetic perturbations on mutation frequency across cell types. These studies will provide an unprecedented systematic map of genes and pathways controlling DNA damage and mutations across different human cell types. In addition, our novel methodology will represent an important asset for the scientific community that can have several biomedical applications in a variety of fields providing an easy tool to study genetic factors that control mutagenesis in health and disease states, including (but not limited to) carcinogenesis.

概括 我们的细胞不断暴露于引起DNA损伤的突变，如果未修复，会导致突变。 在细胞水平上，躯体组织中突变的逐渐积累驱动癌变和 其他疾病。尽管我们对突变过程的理解取得了重大进展，但许多人的答案 基本问题仍然是一个神秘的问题。人体组织突变的遗传原因是什么？如何 人体中的不同细胞类型控制DNA损伤和修复吗？尽管我们的身体很大 在控制模型生物中DNA损伤和修复的途径中的知识，我们缺乏深度 了解人类组织中的这一过程。了解突变如何导致 包括癌变的人类疾病。 在这里，我们将使用一种高度创新的方法来构建Mutsensor，这是一组突变记者来估计 哺乳动物细胞中的DNA突变频率比现有的敏感性高> 50倍 方法。通过“遗传写作和 在Boeke Lab中开发的“交付”系统。这种新颖的方法将允许大规模的功能通用 以前所未有的规模进行筛选。在我们的初步数据的基础上，我们旨在全面确定所有 调节不同人类细胞类型中突变频率的基因。为此，我们将利用损失和 功能收益筛选文库确定遗传扰动对跨越突变频率的影响 细胞类型。 这些研究将为控制DNA损伤的基因和途径提供前所未有的系统图 以及不同人类细胞类型的突变。此外，我们的新方法将代表一个重要的 科学界的资产，可以在各个领域具有几种生物医学应用 提供一个简单的工具来研究控制健康和疾病状态中诱变的遗传因素， 包括（但不限于）癌变。