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基本影响日益被了解。生态失调是一种微生物组细菌成分的不平衡，与多种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组，以治疗和预防此类疾病。该项目将设计益生菌生物来检测具有病理状态特征的代谢变化，并触发在感兴趣的部位部署适当的治疗。这将需要开发益生菌乳酸菌的工程工具，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。我们目前缺乏必要的工具来检测和选择性地去除引起生态失调的微生物组的细菌成分。这限制了治疗干预。新型的下一代治疗细菌（NTB）能够感知和精确解决生态失调，代表了微生物组相关疾病治疗的新前沿。该项目将开发关键的生态失调传感组件，以创造新的细菌疗法，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和反应微生物群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进入微生物组的部署。将采用生物信息学方法来识别控制转录调节以响应外部信号的新传感组件。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一个合成生物学工具箱。对于人类微生物组的治疗用途，乳酸菌（如乳酸杆菌）是理想的选择，因为它们通常被认为是安全的（GRAS）状态。然而，很少有特征启动子、核糖体结合位点、终止子和载体被描述为乳杆菌，因此，解决这一需求将改变我们可靠地设计这些类型的生物体的能力。

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.