CAREER: Programmable synthetic microbial consortia for complex multicellular functions

工程混合细胞群体在生物制造、基于细胞的治疗和诊断方面具有潜在的应用。细菌代谢是一个高活性的网络，不同的反应发生在不同的时间。它们由内部和外部产生的信号打开和关闭。通过这种方式，细胞可以通过特定的反应路径，对碳源和其他材料的流动做出有效的决定。设计细胞群体的尝试并没有达到类似的控制效果。该项目将侧重于为设计微生物混合物的控制系统制定一项协议。该协议将指导构建有助于实现控制系统的基因组件。这些成分将在各种生物体中进行评估。该项目的一个核心目标是教育和指导多样化的未来劳动力。将制定和实施K-12 STEM教育计划和为代表不足的群体拓宽研究机会的计划。与自然发生的动态途径表达类似，计算动态代谢模型预测通过离散代谢状态和临时酶激活来优化生物生产。这在生物生产微生物联盟中是不可能实现的。拟议研究的目标是为细菌联盟的自动化设计建立一个可推广的平台。用户将能够动态控制新陈代谢，并指定对用户定义的时间信号的协调多细胞响应。该项目将开发遗传电路组件和用于在工程细菌群落中划分时序逻辑电路的电路设计算法，并确定合成微生物联合体中生物时序逻辑编程的设计原则。这项拟议的研究将生产与自动化设计算法和用于细菌细胞间信号传递的物理质粒兼容的电路元件。这将使研究人员能够利用现成的部件对在微生物群体中实施时间转录控制的多细胞遗传电路进行先验设计。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Engineered mixed cell populations have potential application in biomanufacturing, cell-based therapies, and diagnostics. Bacterial metabolism is a highly reactive network, and different reactions occur at different times. They are turned on and off by signals generated internally and externally. In this way the cell can make effective decisions regarding the flow of carbon sources and other materials through specific reaction pathways. Attempts to engineer cell populations have not achieved comparable control. This project will focus on developing a protocol for the design of control systems for mixtures of microbes. The protocol will guide the construction of genetic components that can help implement the control system. These components will be evaluated in a variety of organisms. A core objective of this project is to educate and mentor a diverse future workforce. Programs for K-12 STEM education and to broaden research opportunities for underrepresented groups will be developed and implemented.Similar to naturally-occurring dynamic pathway expression, computational dynamic metabolic modeling predicts optimized bioproduction by discrete metabolic states and temporal enzyme activation. This has not been achievable within bioproduction microbial consortia. The goal of the proposed research is to establish a generalizable platform for the automated design of bacterial consortia. Users will be able to dynamically control metabolism and specify coordinated multicellular responses to user-defined temporal signals. This project will develop genetic circuit components and a circuit design algorithm for partitioning sequential logic circuits within engineered bacterial communities, and identify design principles for biological sequential logic programming in synthetic microbial consortia. The proposed research will produce circuit components that are compatible with automated design algorithms and physical plasmids for bacterial intercellular signaling. This will enable researchers to utilize off-the-shelf parts for a priori design of multicellular genetic circuits that implement temporal transcription control in microbial consortia.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.