Clostridioes difficile is a Gram-positive, spore-forming, obligate anaerobic bacterium, and the main cause of hospital-associated diarrhea. In recent years, with the presence of virulent strains (i.e., ribosome type 027), the prevalence and mortality events have increased. Thus, studies on physiological and biochemical characteristics, and pathogenic mechanisms of C. difficile have been performed. The development of efficient and stable genome-editing methods for C. difficile is urgent for the dissection of its physiological and pathogenic mechanism. For example, ClosTron technology plays a key role in study of the relationship between C. difficile toxins (Toxin A and Toxin B) and its pathogenicity. This article reviews the history, recent progress and future prospects of C. difficile genome-editing technologies.
艰难梭菌Clostridioes difficile 是一种革兰氏阳性、产芽孢、专性厌氧细菌，是医院相关性腹泻的主要病原体。近年来，随着强毒株的出现 (如核糖体027 型)，其流行性与致死率逐年上升，因此对艰难梭菌生理、生化特征及致病机制的研究受到广泛重视。艰难梭菌生理、生化特征及致病机制研究又以建立其稳定、高效的基因编辑方法为必要前提。借助基因编辑工具，研究者可以扰动艰难梭菌核心生物学过程，在分子水平研究其分子致病机制。如ClosTron 技术在艰难梭菌毒素A (Toxin A) 和毒素B (Toxin B) 与其致病力关系的研究中起到了关键作用。文中以时间为主线综述了艰难梭菌基因编辑技术的发展历程和最新进展，并对艰难梭菌基因编辑技术未来的研究方向进行展望。.
Keywords: Clostridioes difficile; Clostridioes difficile infection (CDI); gene function research; genome editing; pathogenic mechanism.