High‐efficiency delivery of CRISPR‐Cas9 by engineered probiotics enables precise microbiome editing

Molecular Systems Biology

Alfredo Menendez¹  Frédéric Grenier²  Kevin Neil²  Vincent Burrus²  Nancy Allard²  Sébastien Rodrigue²  Patricia Roy² 

[1] Département de Microbiologie et d'Infectiologie Université de Sherbrooke Sherbrooke QC Canada;Département de biologie Université de Sherbrooke Sherbrooke QC Canada;

关键词: antibiotic resistance;    antimicrobials;    bacterial conjugation;    CRISPR‐Cas9;    synthetic biology;   

DOI  :  10.15252/msb.202110335

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown