Supplement for Purchase of a Cell Sorter for R35GM133797

项目摘要 细胞异质性在生命的所有领域都是普遍存在的。基因相同的细胞可以 即使在相同的环境中生长，也显示出异质的代谢活动。在 细菌，代谢异质性可以塑造整体生长速率，并影响抗生素 宽容在细菌中，代谢异质性塑造了整体生长速率， 抗生素耐受性然而，许多问题仍然关于代谢异质性。为 例如，是什么决定了代谢波动的大小？代谢异质性 监管？我们能否控制代谢异质性，从而消除耐药细胞？ 由于缺乏基本的了解，严重限制了有效的发展。 治疗多种疾病，其中少数短暂耐受细胞经常引起 疾病复发。目前的MIRA项目旨在系统地了解 利用单细胞代谢分析方法研究细菌代谢异质性。使用 工程代谢途径，产生一种独特的荧光代谢物-betaxanthine，这 该项目旨在确定代谢异质性的起源、动力学和传播。 此外，该项目还研究了转录调控如何影响代谢异质性， 并探讨了代谢异质性对药物耐受性的影响，以及降低药物耐受性的策略。 代谢异质性 此设备补充申请要求资金购买新的流式细胞仪， 更换PI实验室中损坏且无法修复的旧流式细胞仪。流式细胞仪 对于研究代谢物异质性的亲本项目至关重要。获取新的流 流式细胞仪将大大加快该项目的生产力，并使新的能力 PI的研究小组探索代谢物异质性的新领域。

PROJECT SUMMARY Cellular heterogeneity is ubiquitous across all domains of life. Genetically identical cells can display heterogeneous metabolic activities, even when grown in identical environments. In bacteria, metabolic heterogeneity can shape the ensemble growth rate and affect antibiotic tolerance. In bacteria, metabolic heterogeneity shapes the ensemble growth rate and affects antibiotic tolerance. However, many questions remain regarding metabolic heterogeneity. For instance, what determines the size of metabolic fluctuation? How is metabolic heterogeneity regulated? Can we control metabolic heterogeneity and therefore eliminate drug-tolerant cells? The lack of fundamental understanding has severely limited the development of effective treatments for multiple diseases in which a small number of transiently tolerant cells often cause disease recurrence. The current MIRA project aims to obtain a systematic understanding of bacteria metabolic heterogeneity by using single-cell metabolic analysis methods. Using an engineered metabolic pathway that produces a unique fluorescent metabolite―betaxanthine, this project aims to determine the origin, dynamics, and propagation of metabolic heterogeneity. Furthermore, this project studies how transcriptional regulations affect metabolic heterogeneity, and explores the influence of metabolic heterogeneity on drug tolerance and strategies to reduce metabolic heterogeneity. This Equipment Supplement application requests fund to purchase a new flow cytometer to replace an old flow cytometer in the PI’s lab that was broken and is not repairable. Flow cytometer is essential to the parental project in studying metabolite heterogeneity. Obtaining a new flow cytometer will substantially accelerate the productivity of this project and enable new capabilities to the PI’s research group to explore new areas in metabolite heterogeneity.