Open AccessHigh‐efficiency delivery of CRISPR‐Cas9 by engineered probiotics enables precise microbiome editing
Author(s) -
Neil Kevin,
Allard Nancy,
Roy Patricia,
Grenier Frédéric,
Menendez Alfredo,
Burrus Vincent,
Rodrigue Sébastien
Publication year - 2021
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.202110335
Subject(s) - crispr , biology , cas9 , plasmid , computational biology , genome editing , antibiotic resistance , microbiome , escherichia coli , microbiology and biotechnology , bacteria , antibiotics , dna , genetics , gene
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.