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J Adv Res. 2019 Jan 31;18:61-69. doi: 10.1016/j.jare.2019.01.011. eCollection 2019 Jul.

Reversal of carbapenem-resistance in Shewanella algae by CRISPR/Cas9 genome editing.

Author information

1
US Department of Energy Joint Genome Institute, Walnut Creek, CA, USA.
2
Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan.
3
Department of Computer Science and Information Engineering, National Chung Cheng University, Chia-Yi, Taiwan.
4
Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan.
5
Division of Infectious Diseases, Department of Internal Medicine, Taichung Veterans General Hospital, Taichung, Taiwan.
6
Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taichung, Taiwan.
7
Department of Physical Therapy, Shu-Zen Junior College of Medicine and Management, Kaohsiung City, Taiwan.
8
Ph.D. Program in Translational Medicine, National Chung Hsing University, Taichung, Taiwan.

Abstract

Antibiotic resistance in pathogens is a growing threat to human health. Of particular concern is resistance to carbapenem, which is an antimicrobial agent listed as critically important by the World Health Organization. With the global spread of carbapenem-resistant organisms, there is an urgent need for new treatment options. Shewanella algae is an emerging pathogen found in marine environments throughout the world that has increasing resistance to carbapenem. The organism is also a possible antibiotic resistance reservoir in humans and in its natural habitat. The development of CRISPR/Cas9-based methods has enabled precise genetic manipulation. A number of attempts have been made to knock out resistance genes in various organisms. The study used a single plasmid containing CRISPR/Cas9 and recE/recT recombinase to reverse an antibiotic-resistant phenotype in S. algae and showed bla OXA-55 -like gene is essential for the carbapenem resistance. This result demonstrates a potential validation strategy for functional genome annotation in S. algae.

KEYWORDS:

Beta-lactamase OXA-55; COGs, Clusters of Orthologous Groups of proteins; CRISPR, clustered regularly interspaced short palindromic repeat; CRISPR-Cas9; Carbapenem; Cas9, CRISPR-associated protein 9; DAP, diaminopimelic acid; NCBI, National Center for Biotechnology Information; PGAAP, Prokaryotic Genomes Automatic Annotation Pipeline; Resistance; SMRT, single-molecule real-time; Shewanella algae

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