EAGER: Enabling Efficient Genome Editing for Advanced Biomanufacturing Applications

EAGER：为先进的生物制造应用实现高效的基因组编辑

• 批准号: 1539359
• 负责人: Kelvin Lee
• 金额: $ 30万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1539359&HistoricalAwards=false
• 关键词: EAGER; Enabling; Efficient; Genome; Editing

This project addresses the challenge of identifying and correcting problems that affect the efficiency of the cell culture system used to produce the majority of therapeutic proteins. Such biomanufacturing using Chinese hamster ovary (CHO) cells is part of a $100 billion industry. CHO cells are difficult to modify using modern genetic engineering tools and are prone to random changes in their genetic material (DNA). This project will examine proteins that repair damaged DNA as a cause of the difficulties in genetic engineering as well as the genetic instability of CHO cells, and promises to generate new insights into the biology of cell lines as well as developing new tools for these cells. Students will be trained in an interdisciplinary setting that will expose them to multiple potential career paths. This project is anticipated to result in more efficient genome editing in Chinese hamster ovary (CHO) cells; performing synthetic biology on these cells is currently problematic, even using CRISPR and other advanced approaches. The hypothesis will be tested that the DNA double strand break repair system (a component of the molecular machinery that is needed to perform genome editing) is flawed, thereby leading to the failure of synthetic biology approaches in these cells. Defects in the DNA double strand break repair system may also be the cause of the problematic genome instability in CHO cells. The goals of the project are 1) to systematically replace each of the DNA double strand break repair genes from CHO with the corresponding version from the Chinese hamster following a priority list based on sequence similarity and function; and 2) after each replacement, to quantitatively evaluate whether the change has had a measureable impact on efficiency of genome editing and on chromosomal stability.

该项目应对识别和纠正影响细胞培养系统用于生产大多数治疗蛋白的效率的问题的挑战。这种使用中国仓鼠卵巢（CHO）细胞的生物制造是1000亿美元行业的一部分。 CHO细胞很难使用现代基因工程工具进行修改，并且容易随机改变其遗传物质（DNA）。 该项目将检查修复DNA的蛋白质，这是基因工程困难以及CHO细胞的遗传不稳定性的原因，并有望为细胞系生物学以及为这些细胞开发新工具而产生新的见解。 学生将在跨学科的环境中接受培训，这将使他们处于多种潜在的职业道路上。预计该项目将在中国仓鼠卵巢（CHO）细胞中进行更有效的基因组编辑。在这些细胞上进行合成生物学，即使使用CRISPR和其他先进方法也是有问题的。 该假设将被检验，即DNA双链断裂修复系统（进行基因组编辑所需的分子机械的组成部分）是有缺陷的，从而导致这些细胞中合成生物学方法的失败。 DNA双链断裂修复系统中的缺陷也可能是CHO细胞中有问题的基因组不稳定性的原因。 该项目的目标是1）系统地，根据序列相似性和功能，使用中国仓鼠的相应版本从CHO中使用CHO的每个DNA双链断修复基因； 2）每次替换后，定量评估变化是否对基因组编辑效率和染色体稳定性产生了可测量的影响。