High‐efficiency delivery of CRISPR‐Cas9 by engineered probiotics enables precise microbiome editing. Issue 10 (19th October 2021)

• Record Type: Journal Article
• Authors: Neil, Kevin; Allard, Nancy; Roy, Patricia; Grenier, Frédéric; Menendez, Alfredo; Burrus, Vincent; Rodrigue, Sébastien
• Is Part Of: Molecular systems biology. Volume 17:Issue 10(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-10-19
• Subjects: antibiotic resistance -- antimicrobials -- bacterial conjugation -- CRISPR‐Cas9 -- synthetic biology; Molecular biology -- Periodicals; Systems biology -- Periodicals; 572.8
• DOI: 10.15252/msb.202110335
• Languages: English
• ISSNs: 1744-4292

Abstract

Abstract: Antibiotic resistance threatens our ability to treat infectious diseases, spurring interest in alternative antimicrobial technologies. The use of bacterial conjugation to deliver CRISPR‐ cas systems programmed to precisely eliminate antibiotic‐resistant bacteria represents a promising approach but requires high in situ DNA transfer rates. We have optimized the transfer efficiency of conjugative plasmid TP114 using accelerated laboratory evolution. We hence generated a potent conjugative delivery vehicle for CRISPR‐ cas9 that can eliminate > 99.9% of targeted antibiotic‐resistant Escherichia coli in the mouse gut microbiota using a single dose. We then applied this system to a Citrobacter rodentium infection model, achieving full clearance within four consecutive days of treatment. SYNOPSIS: A conjugative plasmid with high transfer rates is leveraged to deliver the CRISPR system into targeted bacteria. The resulting conjugative system can clear a C. rodentium infection in mice. Conjugative plasmid TP114 can be used for the delivery of CRISPR‐Cas systems by an engineered conjugative probiotic (COP) strain. Accelerated laboratory evolution was conducted to further increase TP114 transfer rates in the gut microbiota. The COP approach was able to knock down antibiotic‐resistant bacteria from a probed population in situ . The improved eB‐COP system enabled the complete clearance of a C. rodentium infection in mice with similar efficiency as a conventional antibiotic treatment. Abstract : A conjugative plasmid with high transfer rates is leveraged to deliver the CRISPR system into targeted bacteria. The resulting conjugative system can clear a C. rodentium infection in mice.