Application of CRISPR-Cas System to Mitigate Superbug Infections

Abstract

Multidrug resistance in bacterial strains known as superbugs is estimated to cause fatal infections worldwide. Migration and urbanization have resulted in overcrowding and inadequate sanitation, contributing to a high risk of superbug infections within and between different communities. The CRISPR-Cas system, mainly type II, has been projected as a robust tool to precisely edit drug-resistant bacterial genomes to combat antibiotic-resistant bacterial strains effectively. To entirely opt for its potential, advanced development in the CRISPR-Cas system is needed to reduce toxicity and promote efficacy in gene-editing applications. This might involve base-editing techniques used to produce point mutations. These methods employ designed Cas9 variations, such as the adenine base editor (ABE) and the cytidine base editor (CBE), to directly edit single base pairs without causing DSBs. The CBE and ABE could change a target base pair into a different one (for example, G-C to A-T or C-G to A-T). In this review, we addressed the limitations of the CRISPR/Cas system and explored strategies for circumventing these limitations by applying diverse base-editing techniques. Furthermore, we also discussed recent research showcasing the ability of base editors to eliminate drug-resistant microbes.

Recent advances in gene-editing approaches for tackling antibiotic resistance threats: a review

“…Talking about other gene editing/targeting approaches, CRISPR/Cas-based techniques have become a widely adopted, promising complement against conventional approaches to fighting against antibiotic resistance with minimal harmful repercussions on humans or the environment. CRISPR-Cas genome editing was praised for its simplicity, effectiveness, adaptability, and lack of requirement for any markers for recognizing species of harmful bacteria as compared to other DNA-based genetic engineering techniques ( Rabaan et al., 2023 ). Despite this, it can accurately target a particular sequence using just one guide RNA (gRNA) and the protein that accompanies it (Cas).…”

“…A modifying enzyme for precise nucleotide modification and an ssDNA for programmable DNA binding are the two main parts of base editors, that allow this method to change bases properly. DNA base editors use fusion proteins (nickase Cas9, dead Cas9, or dead Cas12a/b) fused to ssDNA-specific nucleobase deaminases to increase the efficacy of site-directed mutagenesis ( Kantor et al., 2020 ; Rabaan et al., 2023 ). Moreover, it is very important to understand that the two gene editing methods that preceded the CRISPR/Cas 9 technology were TALEN and ZFN respectively.…”