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CRISPR/Cas9/sgRNA-mediated targeted gene modification confirms the cause-effect relationship between gyrA mutation and quinolone resistance in Escherichia coli.
FEMS Microbiol Lett. 2018 07 01; 365(13)FM

Abstract

Quinolones are broad-spectrum antibiotics that have been used for decades in treating bacterial infections in humans and animals, and subsequently bacterial resistance to these agents has increased. While studies indicated the relationship between gyrA mutations and bacterial resistance to quinolones, CRISPR/Cas9 was used in this study to investigate causal role of gyrA mutation in the quinolone resistance. In this study, 818 clinical Escherichia coli isolates were analyzed for gyrA mutations and their resistance to quinolones. The CRISPR/Cas9 system was used to generate gyrA mutations in quinolone-susceptible E. coli ATCC 25922, and quinolone-resistant clinical E. coli. The antimicrobial resistance prevalence rate in E. coli against nalidixic acid, ciprofloxacin and enrofloxacin was 77.1% (631/818), 51.1% (418/818) and 49.8% (407/818), respectively. The gyrA mutations were identified in nucleotide positions 248, 255, 259, 260, 261, 273 and 300, and mutations at positions 248 and 259 resulting in amino acid changes at positions 83 and 87 were associated with quinolone resistance. Double-site amino acid mutations increase resistance to quinolones. The gyrA mutations causing changes at amino acids 83 and 87 reversed the features of quinolone resistance in ATCC and clinical strains, verifying the causal role of gyrA mutation in the quinolone resistance of E. coli.

Authors+Show Affiliations

Department of Basic Sciences, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, 225009, PR China.Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, 221003, PR China.Department of Biosciences, Ross University School of Veterinary Medicine, P.O. Box 334, Basseterre, St. Kitts, West Indies. Department of Pathology, Bacteriology and Poultry Diseases, Faculty of Veterinary Medicine Salisburylaan 133, 9820 Merelbeke, Belgium.Department of Basic Sciences, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, 225009, PR China.Department of Basic Sciences, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, 225009, PR China.Department of Basic Sciences, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, 225009, PR China.Department of Basic Sciences, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, 225009, PR China.College of Veterinary Medicine, Auburn University, Auburn, AL, 36849, USA.Department of Basic Sciences, Yangzhou University College of Veterinary Medicine, Yangzhou, Jiangsu, 225009, PR China.College of Veterinary Medicine, Auburn University, Auburn, AL, 36849, USA.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

29767711

Citation

Qiu, Haixiang, et al. "CRISPR/Cas9/sgRNA-mediated Targeted Gene Modification Confirms the Cause-effect Relationship Between gyrA Mutation and Quinolone Resistance in Escherichia Coli." FEMS Microbiology Letters, vol. 365, no. 13, 2018.
Qiu H, Gong J, Butaye P, et al. CRISPR/Cas9/sgRNA-mediated targeted gene modification confirms the cause-effect relationship between gyrA mutation and quinolone resistance in Escherichia coli. FEMS Microbiol Lett. 2018;365(13).
Qiu, H., Gong, J., Butaye, P., Lu, G., Huang, K., Zhu, G., Zhang, J., Hathcock, T., Cheng, D., & Wang, C. (2018). CRISPR/Cas9/sgRNA-mediated targeted gene modification confirms the cause-effect relationship between gyrA mutation and quinolone resistance in Escherichia coli. FEMS Microbiology Letters, 365(13). https://doi.org/10.1093/femsle/fny127
Qiu H, et al. CRISPR/Cas9/sgRNA-mediated Targeted Gene Modification Confirms the Cause-effect Relationship Between gyrA Mutation and Quinolone Resistance in Escherichia Coli. FEMS Microbiol Lett. 2018 07 1;365(13) PubMed PMID: 29767711.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - CRISPR/Cas9/sgRNA-mediated targeted gene modification confirms the cause-effect relationship between gyrA mutation and quinolone resistance in Escherichia coli. AU - Qiu,Haixiang, AU - Gong,Jiansen, AU - Butaye,Patrick, AU - Lu,Guangwu, AU - Huang,Ke, AU - Zhu,Guoqiang, AU - Zhang,Jilei, AU - Hathcock,Terri, AU - Cheng,Darong, AU - Wang,Chengming, PY - 2018/03/27/received PY - 2018/05/11/accepted PY - 2018/5/17/pubmed PY - 2019/9/19/medline PY - 2018/5/17/entrez JF - FEMS microbiology letters JO - FEMS Microbiol Lett VL - 365 IS - 13 N2 - Quinolones are broad-spectrum antibiotics that have been used for decades in treating bacterial infections in humans and animals, and subsequently bacterial resistance to these agents has increased. While studies indicated the relationship between gyrA mutations and bacterial resistance to quinolones, CRISPR/Cas9 was used in this study to investigate causal role of gyrA mutation in the quinolone resistance. In this study, 818 clinical Escherichia coli isolates were analyzed for gyrA mutations and their resistance to quinolones. The CRISPR/Cas9 system was used to generate gyrA mutations in quinolone-susceptible E. coli ATCC 25922, and quinolone-resistant clinical E. coli. The antimicrobial resistance prevalence rate in E. coli against nalidixic acid, ciprofloxacin and enrofloxacin was 77.1% (631/818), 51.1% (418/818) and 49.8% (407/818), respectively. The gyrA mutations were identified in nucleotide positions 248, 255, 259, 260, 261, 273 and 300, and mutations at positions 248 and 259 resulting in amino acid changes at positions 83 and 87 were associated with quinolone resistance. Double-site amino acid mutations increase resistance to quinolones. The gyrA mutations causing changes at amino acids 83 and 87 reversed the features of quinolone resistance in ATCC and clinical strains, verifying the causal role of gyrA mutation in the quinolone resistance of E. coli. SN - 1574-6968 UR - https://www.unboundmedicine.com/medline/citation/29767711/CRISPR/Cas9/sgRNA_mediated_targeted_gene_modification_confirms_the_cause_effect_relationship_between_gyrA_mutation_and_quinolone_resistance_in_Escherichia_coli_ L2 - https://academic.oup.com/femsle/article-lookup/doi/10.1093/femsle/fny127 DB - PRIME DP - Unbound Medicine ER -