工程改造大肠杆菌Nissle 1917以诊断、记录和改善小鼠炎症性肠病的方案。
Protocol for engineering E. coli Nissle 1917 to diagnose, record, and ameliorate inflammatory bowel disease in mice.
作者信息
Zou Zhen-Ping, Fan Yu-Hang, Du Yue, Fang Ting-Ting, Du Wei, Zhou Ying, Ye Bang-Ce
机构信息
Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
出版信息
STAR Protoc. 2023 Apr 26;4(2):102254. doi: 10.1016/j.xpro.2023.102254.
Engineered microorganisms hold potential for disease diagnosis and treatment. Here, we present a protocol to engineer E. coli Nissle 1917 strain (iROBOT) using genome insertion and plasmid construction to diagnose, record, and ameliorate inflammatory bowel disease in mice. We describe steps for constructing and administering iROBOT, diagnosing and recording colitis, preparing samples, and analyzing fluorescence and base editing ratios of iROBOT. We detail a colitis ameliorating assay using the disease activity index, colon length, tissue pathological section, and cytokine analysis. For complete details of the use and execution of this protocol, please refer to Zou et al..
工程微生物在疾病诊断和治疗方面具有潜力。在此，我们展示了一种利用基因组插入和质粒构建来改造大肠杆菌Nissle 1917菌株（iROBOT）的方案，以诊断、记录和改善小鼠的炎症性肠病。我们描述了构建和施用iROBOT、诊断和记录结肠炎、制备样本以及分析iROBOT的荧光和碱基编辑比率的步骤。我们详细介绍了一种使用疾病活动指数、结肠长度、组织病理切片和细胞因子分析的结肠炎改善测定法。有关本方案使用和执行的完整详细信息，请参考邹等人的研究。
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