CAREER: Investigating the DNA Repair Mechanisms of Non-Traditional Genome Editing Agents

职业：研究非传统基因组编辑剂的 DNA 修复机制

• 批准号: 2048207
• 负责人: Alexis Komor
• 金额: $ 130万
• 依托单位: University of California-San Diego
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2048207&HistoricalAwards=false
• 关键词: CAREER; Investigating; DNA; Repair; Mechanisms; CAREER; Investigating; DNA; Repair; Mechanisms

Genome editing tools have driven a revolution and enabled the work of countless researchers across the fields of biomedical science, agriculture, biotechnology, and synthetic biology. Despite the fervor with which these technologies have been adopted, many of them suffer from key shortcomings such as unwanted DNA sequence modifications and low efficiencies for certain types of edits and in certain types of cells. This project seeks to tackle these limitations by furthering basic understanding of DNA repair mechanisms involved in the editing processes. The research will enable expansion of the current suite of genome editing tools, while simultaneously providing graduate student and postdoctoral researchers with training for successful careers in science. The project also includes a research-practice partnership with San Diego area public high schools with high enrollments of socioeconomically disadvantaged students. The students will be introduced to cutting edge genome editing technologies in a topical and approachable manner, with the goal of inciting excitement and further engagement with higher education beyond standard classroom curricula.To alter the sequence of genomic DNA at will, genome editing tools all rely on initial introduction of damage into the DNA of live cells. “Nontraditional” genome editing tools, such as base editors and prime editors, are a class of recently developed editors that employ other types of DNA damage than double-stranded breaks to mediate the editing process. This project will use high-throughput gene knock-down strategies in combination with reporters for various base editing and prime editing outcomes to characterize the role of DNA repair genes in processing nontraditional genome editing intermediates. This effort will be complemented by computational and experimental research to determine how base editing and prime editing outcomes change in relation to different stages of the cell cycle. These studies represent the first investigations into DNA repair mechanisms employed by nontraditional genome editors, and the outcomes will advance the efficiency and precision of existing editing tools as well generate editing tools for new types of genomic modifications.This project is jointly funded by the Genetic Mechanisms and Systems and Synthetic Biology programs of the Molecular and Cellular Biosciences Division in the Biological Sciences DirectorateThis 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.

基因组编辑工具驱动了一场革命，并使在生物医学，农业，生物技术和合成生物学领域的无数研究人员的工作。尽管采用了这些技术的热情，但其中许多人遭受了关键的缺点，例如某些类型的编辑和某些类型的细胞等不需要的DNA序列修改和低效率。该项目试图通过进一步了解编辑过程中涉及的DNA修复机制的基本了解来解决这些局限性。该研究将使当前的基因组编辑工具套件扩展，同时为研究生和博士后研究人员提供科学成功职业的培训。该项目还包括与圣地亚哥地区公立高中的研究实践合作伙伴关系，该公立高中的社会经济不安学生入学率很高。将以局部且平易近人的方式介绍学生以尖锐的基因组编辑技术，以煽动兴奋和进一步与高等教育的接触，而不是标准课堂课程以外的高等教育。要随意改变基因组DNA的顺序，基因组编辑工具都依赖于最初的损害引入活细胞的DNA。 “非传统”基因组编辑工具（例如基础编辑者和主要编辑者）是一类最近开发的编辑器，这些编辑者与双链休息相比其他类型的DNA损伤来介绍编辑过程。该项目将使用高通量基因敲除策略与记者结合使用，用于各种基础编辑和主要编辑结果，以表征DNA修复基因在处理非传统基因组编辑中间体中的作用。计算和实验研究将完成这项工作，以确定基础编辑和主要编辑结果如何与细胞周期的不同阶段有关。这些研究代表了非传统基因组编辑器所携带的DNA修复机制的首次研究，结果将提高现有编辑工具的效率和精确性，并为新型基因组修饰的新类型生成编辑工具，该项目由该项目共同资助了遗传机制和系统生物学生物学分隔的遗传机制和综合生物学分隔中的遗传学生物学分隔，以实现分子生物镜分隔为单位。法定使命，并使用基金会的知识分子优点和更广泛的影响标准通过评估被认为是宝贵的支持。