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Updated: Sep 14, 2025

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira
Published on: August 14, 2021
Precise virulence inactivation using a CRISPR-associated transposase for combating Enterobacteriaceae gut pathogens
Carlotta Ronda1,2, Tyler Perdue1,3, Logan Schwanz1,4
1Department of System Biology, Columbia University, New York, NY, USA.
Abstract:
Targeted gene manipulation in a complex microbial community is an enabling technology for precise microbiome editing. Here we introduce BACTRINS, an in situ microbiome engineering platform designed for efficient and precise genomic insertion of a desired payload and simultaneous knockout of target genes. This system leverages conjugation-mediated delivery of CRISPR-associated transposases to achieve RNA-guided genomic integration, allowing precise insertion of a therapeutic payload while neutralizing pathogen virulence without causing cell death. When applied against an Enterobacteriaceae Shiga toxin-producing pathogen in the gut, this system delivers a CRISPR-associated transposase by bacterial conjugation for site-specific inactivation of the Shiga toxin gene and integration of a nanobody therapeutic payload to disrupt pathogen attachment. A single dose of this therapy results in high-efficiency Shiga gene inactivation and improved survival in a murine infection model of Shiga-producing pathogen. This work establishes a new type of live bacterial therapeutic capable of reducing gut infections by transforming toxigenic pathogens into commensal protectors.
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