☆ 4.0 Article

Zinc Finger Nuclease: A New Approach to Overcome Beta-Lactam Antibiotic Resistance

JUNDISHAPUR JOURNAL OF MICROBIOLOGY (2016)

Journal

JUNDISHAPUR JOURNAL OF MICROBIOLOGY

Volume 9, Issue 1, Pages -

Publisher

AHVAZ JUNDISHAPUR UNIV MED SCI

DOI: 10.5812/jjm.29384

Keywords

Ampicillin Resistance; Microbial Drug Resistance; Beta-Lactamases; Gene Targeting; Genome Editing; Zinc Finger Nuclease

Funding

Isfahan University of Medical Sciences [393358]

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Abstract

Background: The evolution of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs) has been accelerated recently by the indiscriminate application of antibiotics. Antibiotic resistance has challenged the success of medical interventions and therefore is considered a hazardous threat to human health. Objectives: The present study aimed to describe the use of zinc finger nuclease (ZFN) technology to target and disrupt a plasmid-encoded beta-lactamase, which prevents horizontal gene transfer-mediated evolution of ARBs. Materials and Methods: An engineered ZFN was designed to target a specific sequence in the ampicillin resistance gene (amp(R)) of the pTZ57R plasmid. The Escherichia coli bacteria already contained the pZFN kanamycin-resistant (kana(R)) plasmid as the case or the pP15A, kana(R) empty vector as the control, were transformed with the pTZ57R; the ability of the designed ZFN to disrupt the beta-lactamase gene was evaluated with the subsequent disturbed ability of the bacteria to grow on ampicillin (amp) and ampicillin-kanamycin (amp-kana)containing media. The effect of mild hypothermia on the ZFN gene targeting efficiency was also evaluated. Results: The growth of bacteria in the case group on the amp and amp-kana-containing media was significantly lower compared with the control group at 37 degrees C (P < 0.001). Despite being more efficient in hypothermic conditions at 30 degrees C (P < 0.001), there were no significant associations between the incubation temperature and the ZFN gene targeting efficiency. Conclusions: Our findings revealed that the ZFN technology could be employed to overcome ampicillin resistance by the targeted disruption of the ampicillin resistance gene, which leads to inactivation of beta-lactam synthesis. Therefore, ZFN technology could be engaged to decrease the antibiotic resistance issue with the construction of a ZFN archive against different ARGs. To tackle the resistance issue at the environmental level, recombinant phages expressing ZFNs against different ARGs could be constructed and released into both hospital and urban wastewater systems.

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Unclasping potentials of genomics and gene editing in chickpea to fight climate change and global hunger threat

Charul Singh, Ramesh Kumar, Hansa Sehgal, Sharmista Bhati, Tripti Singhal, Gayacharan, M. S. Nimmy, Renu Yadav, Santosh Kumar A. Gupta, Naglaa Abdallah, Aladdin Hamwieh, Rajendra Kumar

Summary: Genomics and genome editing offer great opportunities for crop improvement and fundamental research. The emergence of new genome editing techniques such as zinc finger nucleases (ZFNs), homing endonucleases, transcription activator-like effector nucleases (TALENs), Base Editors (BEs), and Primer Editors (PEs) enables scientists to modulate gene expressions or create novel genes with high precision and efficiency. However, these techniques are expensive and laborious, requiring protein engineering as a prerequisite. Unlike first-generation genome editing methods, CRISPR/Cas9 is easy to construct and can theoretically target multiple locations in the genome with different guide RNAs. This study discusses the progress in genome editing tools, their applications in chickpea crop development, scientific limitations, and future perspectives for enhancing drought resistance, heat tolerance, and yield in chickpea to address global climate change, hunger, and nutritional threats.

FRONTIERS IN GENETICS (2023)