Function of the CRISPR-Cas System of the Human Pathogen Clostridium difficile

MBio
Pierre BoudryOlga Soutourina

Abstract

Clostridium difficile is the cause of most frequently occurring nosocomial diarrhea worldwide. As an enteropathogen, C. difficile must be exposed to multiple exogenous genetic elements in bacteriophage-rich gut communities. CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems allow bacteria to adapt to foreign genetic invaders. Our recent data revealed active expression and processing of CRISPR RNAs from multiple type I-B CRISPR arrays in C. difficile reference strain 630. Here, we demonstrate active expression of CRISPR arrays in strain R20291, an epidemic C. difficile strain. Through genome sequencing and host range analysis of several new C. difficile phages and plasmid conjugation experiments, we provide evidence of defensive function of the CRISPR-Cas system in both C. difficile strains. We further demonstrate that C. difficile Cas proteins are capable of interference in a heterologous host, Escherichia coli. These data set the stage for mechanistic and physiological analyses of CRISPR-Cas-mediated interactions of important global human pathogen with its genetic parasites. Clostridium difficile is the major cause of nosocomial infections associated with antibiotic therapy world...Continue Reading

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Citations

Nov 19, 2017·Applied and Environmental Microbiology·Julian R GarneauLouis-Charles Fortier
May 9, 2019·Toxins·Olga Soutourina
May 16, 2019·Journal of Bacteriology·Kathleen N McAllister, Joseph A Sorg
Aug 16, 2018·Frontiers in Microbiology·Anna MaikovaOlga Soutourina
Jan 23, 2018·Frontiers in Microbiology·Chinmay V Tikhe, Claudia Husseneder
Apr 26, 2019·The CRISPR Journal·Jake L WeissmanPhilip L F Johnson
Aug 11, 2019·Applied and Environmental Microbiology·Anna MaikovaOlga Soutourina
Sep 21, 2018·Frontiers in Microbiology·Louis-Charles Fortier
Feb 14, 2021·FEMS Microbiology Reviews·Joshua HeulerXingmin Sun
May 1, 2021·Science·Ming LiHua Xiang
Jun 3, 2021·International Journal of Molecular Sciences·Shakhinur Islam MondalColin Hill

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Methods Mentioned

BETA
PCR
RNA-seq
phage sequencing

Software Mentioned

CLUSTALW
Mfold
CRISPRTarget
WebLogo
PAM WebLogo
BLASTN
CRISPRdb
Bowtie

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