Protocol for engineering E. coli Nissle 1917 to diagnose, record, and ameliorate inflammatory bowel disease in mice

Zhen-Ping Zou; Yu-Hang Fan; Yue Du; Ting-Ting Fang; Wei Du; Ying Zhou; Bang-Ce Ye

doi:10.1016/j.xpro.2023.102254

Protocol for engineering E. coli Nissle 1917 to diagnose, record, and ameliorate inflammatory bowel disease in mice

STAR Protoc. 2023 Apr 26;4(2):102254. doi: 10.1016/j.xpro.2023.102254. Online ahead of print.

Authors

Zhen-Ping Zou¹, Yu-Hang Fan¹, Yue Du¹, Ting-Ting Fang¹, Wei Du¹, Ying Zhou², Bang-Ce Ye³

Affiliations

¹ Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
² Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China. Electronic address: zhouying@ecust.edu.cn.
³ Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China; Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, China. Electronic address: bcye@ecust.edu.cn.

Abstract

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.¹.

Keywords: Biotechnology and Bioengineering; CRISPR; Health Sciences; Microbiology; Model Organisms; Molecular Biology.