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

人们越来越了解以微生物群为基础的细菌对人类健康的根本影响。微生物失调是一种微生物群细菌成分的失衡，与一系列疾病的病理密切相关。该项目将开发新的方法来重新平衡微生物群，以治疗和预防此类疾病。该项目将对益生菌进行改造，以检测具有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。这将需要开发益生菌乳酸菌的工程工具，以及识别创建所需生物传感器所需的新传感组件的生物信息学方法。目前，我们缺乏必要的工具来检测和选择性地去除导致微生物组失调的细菌成分。这限制了治疗干预。新型的下一代治疗性细菌(NTB)能够感知和精确地解决生态失调，是治疗微生物组相关疾病的新前沿。该项目将开发关键的微生物失调传感组件，通过创建一个生物传感工具箱，使乳酸菌的基因工程能够感知微生物组失调的代谢变化并做出反应，从而创造新的细菌疗法。最终，这些细菌生物传感器将控制NTB将特定杀菌有效载荷部署到微生物组中。我们将采用生物信息学的方法来识别新的传感组件，这些组件管理对外部信号的转录调控。然后，这些将被重新用作生物传感器，用于检测微生物组的状态。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。乳酸菌等乳酸菌在人体微生物群中的治疗用途。考虑到它们通常被认为是安全的(GRAS)状态，这是一个理想的选择。然而，鲜有乳杆菌特有的启动子、核糖体结合位点、终止子和载体被描述，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.