Mechanisms of CRISPR-Cas9-Mediated Gene Regulation in Pseudomonas aeruginosa PAO1
DOI:
https://doi.org/10.70749/ijbr.v4i2.2915Keywords:
CRISPR-Cas9, Pseudomonas aeruginosa PAO1, Antimicrobial Resistance, Genome Editing, CRISPRi, Gene Regulation, Multidrug Resistance.Abstract
The genome editing tool, known as CRISPR-Cas9, is a powerful tool that can be employed in the fight against the multidrug-resistant opportunistic pathogen, Pseudomonas aeruginosa PAO1, which is known for causing severe infections. This review will explore the use of the CRISPR-Cas9 system in learning more about the bacterium and coming up with new ways of fighting infections. We will also look at the basic components of the CRISPR-Cas9 system, which include the Cas9 nuclease activity, specificity of the RNA, and the DNA repair machinery, which includes both homologous recombination and non-homologous end joining. We will also look at the different components of the system in the context of bacteria. This review also explores the different variants of the system, which include the catalytically dead Cas9 (dCas9) for gene silencing and the use of the system for overexpressing specific genes, known as the CRISPR activation (CRISPRa) system. This can be employed in the context of altering specific genes involved in antibiotic resistance, efflux pumps, biofilm, and virulence. We also discuss how effective it is in reversing antimicrobial resistance by targeting and degrading the genes for antimicrobial resistance, eliminating the plasmids, and making the drug-resistant strains susceptible to normal antibiotics. We also discuss the new applications for epigenetic modifications, genome-wide functional screening, and the fusion of these with phage therapy and nanoparticles. Although there have been many improvements in the field, there are still challenges in finding the most effective drug delivery methods, reducing side effects, and the ability of P. aeruginosa to alter its genes. Future perspectives include combining CRISPR-Cas9 with other cutting-edge technologies to develop synergistic approaches for combating this resilient pathogen and solving the antimicrobial resistance crisis.
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