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携带CRISPR-Cas的压力。

The stress of carrying CRISPR-Cas.

作者信息

Haider Dorina, Bauer Richard, Grempels Aline, Roscher Rebecca, Aslan Cagil Ceren, Mauerer Stefanie, Spellerberg Barbara

机构信息

Institute of Medical Microbiology and Hygiene, University Hospital Ulm, Ulm, Germany.

出版信息

Virulence. 2025 Dec;16(1):2541701. doi: 10.1080/21505594.2025.2541701. Epub 2025 Aug 4.

Abstract

() is a commensal that can cause severe invasive bacterial infections. A considerable percentage of strains harbour CRISPR-Cas systems, which apart from being a bacterial immunity system, can play an important role in adaptation to environmental stress. The functionality of CRISPR-Cas systems has not been investigated previously. To address this, we created a set of deletion mutants in the CRISPR-Cas type II-A system of the SK52 type strain, targeting the nuclease Cas9 and CRISPR array. We were able to confirm CRISPR-Cas activity by testing these strains in a plasmid clearance assay. Furthermore, the impact of the CRISPR-Cas system on the viability of was investigated under various stress conditions, such as UV light, hydrogen peroxide exposure, and high temperatures in wild-type and CRISPR-Cas mutant strains. Under these conditions, survival was significantly lower in the strains carrying . Bacterial growth and metabolic activity in Alamar Blue assays were also negatively affected by the presence of in . In summary, we found that the presence of a functional CRISPR-Cas system in leads to measurable metabolic and fitness costs in the wild-type strain. Carrying was associated with an impaired stress response in our experiments and may explain why many strains of this species lack CRISPR-Cas.

摘要

(某细菌)是一种共生菌,可引起严重的侵袭性细菌感染。相当比例的该细菌菌株含有CRISPR-Cas系统,该系统除作为细菌免疫系统外,在适应环境压力方面也可发挥重要作用。此前尚未对该细菌的CRISPR-Cas系统功能进行研究。为解决这一问题,我们在SK52型菌株的CRISPR-Cas II-A型系统中创建了一组缺失突变体,靶向核酸酶Cas9和CRISPR阵列。我们通过在质粒清除试验中测试这些菌株,证实了CRISPR-Cas活性。此外,还在野生型该细菌和CRISPR-Cas突变株中,研究了CRISPR-Cas系统在紫外线、过氧化氢暴露和高温等各种应激条件下对该细菌生存能力的影响。在这些条件下,携带(相关基因或元件)的菌株存活率显著降低。Alamar Blue试验中的细菌生长和代谢活性也受到该细菌中(相关基因或元件)存在的负面影响。总之,我们发现该细菌中功能性CRISPR-Cas系统的存在会导致野生型菌株出现可测量的代谢和适应性代价。在我们的实验中,携带(相关基因或元件)与应激反应受损有关,这可能解释了为什么该物种的许多菌株缺乏CRISPR-Cas。

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