CRISPR-AMRtracker: A novel toolkit to monitor the antimicrobial resistance gene transfer in fecal microbiota

IF 15.8 1区 医学 Q1 PHARMACOLOGY & PHARMACY Drug Resistance Updates Pub Date : 2024-08-24 DOI:10.1016/j.drup.2024.101142
Gong Li , Teng-Fei Long , Shi-Ying Zhou , Li-Juan Xia , Ang Gao , Lei Wan , Xiao-Yuan Diao , Yu-Zhang He , Ruan-Yang Sun , Jin-Tao Yang , Sheng-Qiu Tang , Hao Ren , Liang-Xing Fang , Xiao-Ping Liao , Ya-Hong Liu , Liang Chen , Jian Sun
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Abstract

The spread of antibiotic resistance genes (ARGs), particularly those carried on plasmids, poses a major risk to global health. However, the extent and frequency of ARGs transfer in microbial communities among human, animal, and environmental sectors is not well understood due to a lack of effective tracking tools. We have developed a novel fluorescent tracing tool, CRISPR-AMRtracker, to study ARG transfer. It combines CRISPR/Cas9 fluorescence tagging, fluorescence-activated cell sorting, 16S rRNA gene sequencing, and microbial community analysis. CRISPR-AMRtracker integrates a fluorescent tag immediately downstream of ARGs, enabling the tracking of ARG transfer without compromising the host cell's antibiotic susceptibility, fitness, conjugation, and transposition. Notably, our experiments demonstrate that sfGFP-tagged plasmid-borne mcr-1 can transfer across diverse bacterial species within fecal samples. This innovative approach holds the potential to illuminate the dynamics of ARG dissemination and provide valuable insights to shape effective strategies in mitigating the escalating threat of antibiotic resistance.

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CRISPR-AMRtracker:监测粪便微生物群中抗菌药耐药性基因转移的新型工具包
抗生素耐药基因(ARGs)的传播,尤其是质粒上携带的抗生素耐药基因的传播,对全球健康构成了重大威胁。然而,由于缺乏有效的追踪工具,人们对 ARGs 在人类、动物和环境微生物群落中的传播范围和频率还不甚了解。我们开发了一种新型荧光追踪工具--CRISPR-AMRtracker,用于研究 ARG 的转移。它结合了 CRISPR/Cas9 荧光标记、荧光激活细胞分拣、16S rRNA 基因测序和微生物群落分析。CRISPR-AMRtracker 将荧光标签整合到 ARGs 的下游,从而能够在不影响宿主细胞的抗生素敏感性、适应性、连接和转座的情况下跟踪 ARG 的转移。值得注意的是,我们的实验证明了 sfGFP 标记的质粒携带的 mcr-1 可以在粪便样本中的不同细菌物种间转移。这种创新方法有望揭示 ARG 的传播动态,并为制定有效策略以缓解不断升级的抗生素耐药性威胁提供有价值的见解。
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来源期刊
Drug Resistance Updates
Drug Resistance Updates 医学-药学
CiteScore
26.20
自引率
11.90%
发文量
32
审稿时长
29 days
期刊介绍: Drug Resistance Updates serves as a platform for publishing original research, commentary, and expert reviews on significant advancements in drug resistance related to infectious diseases and cancer. It encompasses diverse disciplines such as molecular biology, biochemistry, cell biology, pharmacology, microbiology, preclinical therapeutics, oncology, and clinical medicine. The journal addresses both basic research and clinical aspects of drug resistance, providing insights into novel drugs and strategies to overcome resistance. Original research articles are welcomed, and review articles are authored by leaders in the field by invitation. Articles are written by leaders in the field, in response to an invitation from the Editors, and are peer-reviewed prior to publication. Articles are clear, readable, and up-to-date, suitable for a multidisciplinary readership and include schematic diagrams and other illustrations conveying the major points of the article. The goal is to highlight recent areas of growth and put them in perspective. *Expert reviews in clinical and basic drug resistance research in oncology and infectious disease *Describes emerging technologies and therapies, particularly those that overcome drug resistance *Emphasises common themes in microbial and cancer research
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