CRISPR/Cas9-Mediated Re-Sensitization of Antibiotic-Resistant Escherichia coli Harboring Extended-Spectrum β-Lactamases.
Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (CRISPR/Cas9) system, a genome editing technology, was shown to be versatile in treating several antibiotic-resistant bacteria. In the present study, we applied the CRISPR/ Cas9 technology to kill extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli. ESBL bacteria are mostly multidrug resistant (MDR), and have plasmid-mediated antibiotic resistance genes that can be easily transferred to other members of the bacterial community by horizontal gene transfer. To restore sensitivity to antibiotics in these bacteria, we searched for a CRISPR/Cas9 target sequence that was conserved among >1,000 ESBL mutants. There was only one target sequence for each TEM- and SHV-type ESBL, with each of these sequences found in ~200 ESBL strains of each type. Furthermore, we showed that these target sequences can be exploited to re-sensitize MDR cells in which resistance is mediated by genes that are not the target of the CRISPR/Cas9 system, but by genes that are present on the same plasmid as target genes. We believe our Re-Sensitization to Antibiotics from Resistance (ReSAFR) technology, which enhances the practical value of the CRISPR/Cas9 system, will be an effective method of treatment against plasmid-carrying MDR bacteria.
Department of Genetic Engineering and Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon 16419, Republic of Korea.|Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Jun-Seob
JS
Kim
Department of Genetic Engineering and Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon 16419, Republic of Korea.
Da-Hyeong
DH
Cho
Department of Genetic Engineering and Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon 16419, Republic of Korea.
Myeongseo
M
Park
Department of Genetic Engineering and Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon 16419, Republic of Korea.
Woo-Jae
WJ
Chung
Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon 16419, Republic of Korea.
Dongwoo
D
Shin
Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon 16419, Republic of Korea.
Kwan Soo
KS
Ko
Department of Genetic Engineering and Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon 16419, Republic of Korea.
Dae-Hyuk
DH
Kweon
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D064113:CRISPR-Cas Systems / Q000235:genetics*; D024901:Drug Resistance, Multiple, Bacterial; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D019450:Genes, MDR; D010957:Plasmids / Q000235:genetics; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000096:biosynthesis* / Q000235:genetics
D000900:Anti-Bacterial Agents; D001618:beta-Lactamases
Antibiotic resistance; CRISPR/Cas9; Extended spectrum beta-lactamases (ESBLs); Multi-drug resistance (MDR); Re-sensitization