Precision engineering of the probiotic <i>Escherichia coli</i> Nissle 1917 with prime editing.

ABSTRACT: CRISPR-Cas systems are transforming precision medicine with engineered probiotics as next-generation diagnostics and therapeutics. To promote human health and treat disease, engineering probiotic bacteria demands maximal versatility to enable non-natural functionalities while minimizing undesired genomic interferences. Here, we present a streamlined prime editing approach tailored for probiotic Escherichia coli Nissle 1917 utilizing only essential genetic modules, including Cas9 nickase from Streptococcus pyogenes, a codon-optimized reverse transcriptase, and a prime editing guide RNA, and an optimized workflow with longer induction. As a result, we achieved all types of prime editing in every individual round of experiments with efficiencies of 25.0%, 52.0%, and 66.7% for DN

SUBMITTER: Chen P-R 

PROVIDER: S-EPMC11837520 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature