Establishing a synthetic biology toolbox for biosensor development in important probiotic host organisms.

基于微生物组的细菌对人类健康的根本影响越来越被理解。菌群失调是微生物组细菌成分的不平衡，其与多种病症的病理学密切相关。该项目将开发新的方法来重新平衡微生物组，以治疗和预防此类疾病。该项目将设计益生菌生物体，以检测病理状态特征的代谢变化，并在感兴趣的部位触发适当治疗的部署。这将需要开发益生乳酸菌的工程工具，以及生物信息学方法来识别创建所需生物传感器所需的新传感组件。我们目前缺乏检测和选择性去除导致微生态失调的微生物组中细菌成分所需的工具。这限制了治疗干预。能够感测和精确解决生态失调的新型下一代治疗细菌（NTB）代表了治疗微生物组相关疾病的新前沿。该项目将开发关键的生态失调传感组件，通过创建一个生物传感工具箱来创建新的细菌疗法，使乳酸菌的基因工程能够感知和反应微生物组生态失调的代谢变化特征。最终，这些细菌生物传感器将通过NTB控制特定杀菌有效载荷在微生物组中的部署。生物信息学方法将被用来识别新的传感组件，这些传感组件控制响应外部信号的转录调控。然后，这些将被重新用作检测微生物组状态的生物传感器。为了更好地设计益生菌，将为合适的宿主创建合成生物学工具箱。对于在人类微生物组中的治疗用途，乳酸菌如乳杆菌属（Lactobacillus spp.）是一个理想的选择，因为他们一般被视为安全（GRAS）的地位。然而，很少描述乳杆菌属的特征性启动子、核糖体结合位点、终止子和载体，因此，满足这一需求将改变我们可靠地设计这些类型生物的能力。

The fundamental impact of microbiome-based bacteria upon human health is increasingly understood. Dysbiosis is an imbalance of microbiome bacterial constituents that is strongly linked to the pathology of a wide range of conditions. This project will develop new approaches to rebalance the microbiome to treat and prevent such diseases. This project will engineer probiotic organisms to detect metabolic changes that are characteristic of pathologic states and trigger the deployment of appropriate treatments at the site of interest. This will require the development of engineering tools for probiotic lactic acid bacteria, as well as bioinformatics approaches to identify new sensing components required to create the required biosensors.We currently lack the tools necessary to detect and selectively remove bacterial constituents of the microbiome that cause dysbiosis. This limits therapeutic intervention. Novel, next-generation therapeutic bacteria (NTB) capable of sensing and precisely resolving dysbiosis represent a new frontier in the treatment of microbiome related disorders. This project will develop key dysbiosis-sensing components to create new bacterial therapies, by creating a biosensing toolbox enabling genetic engineering of lactic acid bacteria that can sense and react to metabolic changes characteristic of microbiome dysbiosis. Ultimately, these bacterial biosensors will control the deployment of specific bactericidal payloads into the microbiome by NTB.A bioinformatics approach will be taken to identify new sensing components that govern transcriptional regulation in response to external signals. These will then be repurposed as biosensors for the detection of the state of the microbiome. To better be able to engineer probiotic bacteria, a synthetic biology toolbox will be created for suitable hosts. For therapeutic use in the human microbiome, lactic acid bacteria such as Lactobacillus spp. are an ideal choice given their Generally Regarded As Safe (GRAS) status. However, few characterised promoters, ribosome binding sites, terminators and vectors are described for Lactobacillus spp., addressing this need will therefore transform our ability to reliably engineer these types of organism.