Resensitizing Tigecycline- and Colistin-resistant Using an Engineered Conjugative CRISPR/Cas9 System

Overview

Journal Microbiol Spectr

Specialty Microbiology

Date 2024 Feb 22

PMID 38385691

Authors

Haijie Zhang

Bo Chen

Zeyu Wang

Kai Peng

Yuan Liu

Zhiqiang Wang

Affiliations

Soon will be listed here.

Abstract

Importance: The emergence of plasmid-encoded (X4) and isolated from human and animal sources has affected the treatment of tigecycline and colistin, and has posed a significant threat to public health. Tigecycline and colistin are considered as the "last line of defense" for the treatment of multidrug-resistant (MDR) Gram-negative bacterial infections, so there is an urgent need to find a method that can resensitize (X4)-mediated tigecycline-resistant and -mediated colistin-resistant bacteria. In this study, we developed a glutamate-based, chromosomal, plasmid-balanced lethal conjugative CRISPR/Cas9 system, which can simultaneously resensitize (X4)-mediated tigecycline-resistant and -mediated colistin-resistant . The counts of tigecycline- and colistin-resistant bacteria decreased to 1% after the resensitization system was administered. This study opens up new pathways for the development of CRISPR-based tools for selective bacterial pathogen elimination and precise microbiome composition change.

Citing Articles

Developing a Versatile Arsenal: Novel Antimicrobials as Offensive Tools Against Pathogenic Bacteria.

Ma J, Lu Z Microorganisms. 2025; 13(1).

PMID: 39858940 PMC: 11767912. DOI: 10.3390/microorganisms13010172.

Design, potential and limitations of conjugation-based antibacterial strategies.

Derollez E, Lesterlin C, Bigot S Microb Biotechnol. 2024; 17(11):e70050.

PMID: 39548711 PMC: 11568246. DOI: 10.1111/1751-7915.70050.

References

Terns M, Terns R . CRISPR-based adaptive immune systems. Curr Opin Microbiol. 2011; 14(3):321-7. PMC: 3119747. DOI: 10.1016/j.mib.2011.03.005. View

Shenoy E, Macy E, Rowe T, Blumenthal K . Evaluation and Management of Penicillin Allergy: A Review. JAMA. 2019; 321(2):188-199. DOI: 10.1001/jama.2018.19283. View

. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Lancet. 2022; 399(10325):629-655. PMC: 8841637. DOI: 10.1016/S0140-6736(21)02724-0. View

Zheng X, Zhu J, Zhang J, Cai P, Sun Y, Chang M . A novel plasmid-borne tet(X6) variant co-existing with blaNDM-1 and blaOXA-58 in a chicken Acinetobacter baumannii isolate. J Antimicrob Chemother. 2020; 75(11):3397-3399. DOI: 10.1093/jac/dkaa342. View

Wan P, Cui S, Ma Z, Chen L, Li X, Zhao R . Reversal of -Mediated Colistin Resistance in by CRISPR-Cas9 System. Infect Drug Resist. 2020; 13:1171-1178. PMC: 7184118. DOI: 10.2147/IDR.S244885. View

Dorado-Morales P, Lamberioux M, Mazel D . Unlocking the potential of microbiome editing: A review of conjugation-based delivery. Mol Microbiol. 2023; 122(3):273-283. DOI: 10.1111/mmi.15147. View

Rodrigues M, McBride S, Hullahalli K, Palmer K, Duerkop B . Conjugative Delivery of CRISPR-Cas9 for the Selective Depletion of Antibiotic-Resistant Enterococci. Antimicrob Agents Chemother. 2019; 63(11). PMC: 6811441. DOI: 10.1128/AAC.01454-19. View

Wang P, He D, Li B, Guo Y, Wang W, Luo X . Eliminating mcr-1-harbouring plasmids in clinical isolates using the CRISPR/Cas9 system. J Antimicrob Chemother. 2019; 74(9):2559-2565. DOI: 10.1093/jac/dkz246. View

He T, Wang R, Liu D, Walsh T, Zhang R, Lv Y . Emergence of plasmid-mediated high-level tigecycline resistance genes in animals and humans. Nat Microbiol. 2019; 4(9):1450-1456. DOI: 10.1038/s41564-019-0445-2. View

10.

He Y, Yan J, He B, Ren H, Kuang X, Long T . A Transposon-Associated CRISPR/Cas9 System Specifically Eliminates both Chromosomal and Plasmid-Borne in Escherichia coli. Antimicrob Agents Chemother. 2021; 65(10):e0105421. PMC: 8448152. DOI: 10.1128/AAC.01054-21. View

11.

Chung C, Niemela S, Miller R . One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci U S A. 1989; 86(7):2172-5. PMC: 286873. DOI: 10.1073/pnas.86.7.2172. View

12.

Makarova K, Haft D, Barrangou R, Brouns S, Charpentier E, Horvath P . Evolution and classification of the CRISPR-Cas systems. Nat Rev Microbiol. 2011; 9(6):467-77. PMC: 3380444. DOI: 10.1038/nrmicro2577. View

13.

Liu Y, Wang Y, Walsh T, Yi L, Zhang R, Spencer J . Emergence of plasmid-mediated colistin resistance mechanism MCR-1 in animals and human beings in China: a microbiological and molecular biological study. Lancet Infect Dis. 2015; 16(2):161-8. DOI: 10.1016/S1473-3099(15)00424-7. View

14.

Wongpayak P, Meesungnoen O, Saejang S, Subsoontorn P . A highly effective and self-transmissible CRISPR antimicrobial for elimination of target plasmids without antibiotic selection. PeerJ. 2021; 9:e11996. PMC: 8428258. DOI: 10.7717/peerj.11996. View

15.

Karakonstantis S, Kritsotakis E, Gikas A . Treatment options for K. pneumoniae, P. aeruginosa and A. baumannii co-resistant to carbapenems, aminoglycosides, polymyxins and tigecycline: an approach based on the mechanisms of resistance to carbapenems. Infection. 2020; 48(6):835-851. PMC: 7461763. DOI: 10.1007/s15010-020-01520-6. View

16.

Medjitna T, Stadler C, Bruckner L, Griot C, Ottiger H . DNA vaccines: safety aspect assessment and regulation. Dev Biol (Basel). 2006; 126:261-70. View

17.

Lewis K . The Science of Antibiotic Discovery. Cell. 2020; 181(1):29-45. DOI: 10.1016/j.cell.2020.02.056. View

18.

Ryan E, Crean T, Kochi S, John M, Luciano A, Killeen K . Development of a DeltaglnA balanced lethal plasmid system for expression of heterologous antigens by attenuated vaccine vector strains of Vibrio cholerae. Infect Immun. 1999; 68(1):221-6. PMC: 97124. DOI: 10.1128/IAI.68.1.221-226.2000. View

19.

Sun J, Chen C, Cui C, Zhang Y, Liu X, Cui Z . Plasmid-encoded tet(X) genes that confer high-level tigecycline resistance in Escherichia coli. Nat Microbiol. 2019; 4(9):1457-1464. PMC: 6707864. DOI: 10.1038/s41564-019-0496-4. View

20.

Hwang I, Koh E, Wong A, March J, Bentley W, Lee Y . Engineered probiotic Escherichia coli can eliminate and prevent Pseudomonas aeruginosa gut infection in animal models. Nat Commun. 2017; 8:15028. PMC: 5394271. DOI: 10.1038/ncomms15028. View