Targeted gene manipulation in a complex microbial community is an enabling technology for precise microbiome editing. This thesis introduces a new microbial therapeutic system dubbed Bacterial CRISPR-Transposase Reduction of Virulence In Situ (BACTRINS). BACTRINS is an in-situ microbiome engineering platform designed for efficient and precise genomic insertion of a desired payload and simultaneous knockout of target genes. When applied against a 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 resulted 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.
| Identifer | oai:union.ndltd.org:columbia.edu/oai:academiccommons.columbia.edu:10.7916/fvvb-6y70 |
| Date | January 2024 |
| Creators | Perdue, Tyler David |
| Source Sets | Columbia University |
| Language | English |
| Detected Language | English |
| Type | Theses |
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