Using an endogenous CRISPR-Cas system for genome editing in the human pathogen Clostridium difficile

Anna Maikova, Victor Kreis, Anaïs Boutserin, Konstantin Severinov, Olga Soutourina

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    18 Citations (Scopus)


    The human enteropathogen Clostridium difficile constitutes a key public health issue in industrialized countries. Many aspects of C. difficile pathophysiology and adaptation inside the host remain poorly understood. We have recently reported that this bacterium possesses an active CRISPR-Cas system of subtype I-B for defense against phages and other mobile genetic elements that could contribute to its success during infection. In this paper, we demonstrate that redirecting this endogenous CRISPR-Cas system toward autoimmunity allows efficient genome editing in C. difficile. We provide a detailed description of this newly developed approach and show, as a proof of principle, its efficient application for deletion of a specific gene in reference strain 630Δerm and in epidemic C. difficile strain R20291. The new method expands the arsenal of the currently limiting set of gene engineering tools available for investigation of C. difficile and may serve as the basis for new strategies to control C. difficile infections.

    Original languageEnglish
    Article numbere01416-19
    JournalApplied and Environmental Microbiology
    Issue number20
    Publication statusPublished - 2019


    • Clostridium difficile
    • CRISPR
    • Endogenous subtype I-B CRISPR-Cas system
    • Genome editing


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