[3PS-100]
Production of a Probiotic Strain via CRISPR-Based Targeted Gene Editing
발표자윤별 (경상국립대학교)
연구책임자강유경 (경상국립대학교)
Abstract
Escherichia coli Nissle 1917 is a representative probiotic strain that efficiently colonizes the gut and exhibits beneficial functions, including immune modulation, anti-inflammatory, and antimicrobial activities. Due to these properties, EcN has garnered attention as a potential therapeutic for intestinal diseases such as inflammatory bowel disease, Crohn’s disease, ulcerative colitis, and colorectal cancer. However, its clinical translation is hindered by safety concerns regarding the pks gene cluster, which is linked to the genotoxic metabolite colibactin. In this study, we engineered a safety-enhanced EcN strain by deleting a gene involved in colibactin activation using a CRISPR–Cas9–based editing strategy with homologous recombination. The edited candidates were obtained and subsequently stabilized through plasmid curing. This work presents a practical approach to genetically block colibactin-associated risks while preserving EcN’s probiotic potential.