Collaborative Research: Data-driven engineering of the yeast Kluyveromyces marxianus for enhanced protein secretion

合作研究：马克斯克鲁维酵母的数据驱动工程，以增强蛋白质分泌

• 批准号: 2323983
• 负责人: Nancy Da Silva
• 金额: $ 35万
• 依托单位: University of California-Irvine
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-15 至 2027-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2323983&HistoricalAwards=false
• 关键词: Collaborative; Research; Data; driven; engineering

Purification is the limiting step in protein production. If the protein can be secreted out of the cell, purification is much easier and simpler. Certain yeasts are proficient at protein secretion. The objective of this project is to develop a yeast strain with a robust protein secretion system that is also tolerant to extreme pH and temperature levels. Gene editing tools such as CRISPR will enable the improvement of a native yeast. Production of a broad range of proteins will be evaluated. This project also focuses on improving STEM education for underrepresented minorities and non-traditional students at the high school, community college, and undergraduate levels. Improving success in college and the diversity of the US biotechnology workforce are desired outcomes.The secretory pathway of the yeast Kluyveromyces marxianus will be engineered. Genome-wide CRISPR-Cas9 knockout, CRISPRi, and CRISPRa libraries will be used to identify critical genes influencing protein production and secretion. These screens should generate large datasets that link phenotype-to-genotype. This data will be used to inform a second phase of the project. In the second phase, multiplexed CRISPR-based gene-regulation will identify synergistic combinations leading to more efficient protein secretion. The research could result in effective CRISPR gRNA libraries, optimized selection strategies, fundamental knowledge of the secretory pathway in this emerging industrial yeast, and a library of gene interventions leading to substantially higher protein secretion for a range of proteins.This 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.

纯化是蛋白质产生的限制步骤。如果可以从细胞中分泌蛋白质，则纯化更容易，更简单。某些酵母精通蛋白质分泌。该项目的目的是开发具有强大的蛋白质分泌系统的酵母菌菌株，该系统也耐受性pH和温度水平。 CRISPR等基因编辑工具将使原生酵母改善。将评估广泛的蛋白质的生产。该项目还着重于改善高中，社区学院和本科生的代表性不足的少数民族和非传统学生的STEM教育。在大学的成功和美国生物技术劳动力的多样性是理想的结果。全基因组CRISPR-CAS9淘汰，CRISPRI和CRISPRA库将用于识别影响蛋白质生产和分泌的关键基因。这些屏幕应生成将表型与基因型联系起来的大型数据集。该数据将用于告知项目的第二阶段。在第二阶段，基于CRISPR的基因调节将确定协同组合，从而导致更有效的蛋白质分泌。 The research could result in effective CRISPR gRNA libraries, optimized selection strategies, fundamental knowledge of the secretory pathway in this emerging industrial yeast, and a library of gene interventions leading to substantially higher protein secretion for a range of proteins.This 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.