Engineering CRISPR-Cas13 Armed Bacteriophages to Combat Carbapenem Resistant Klebsiella pneumonia: A Targeted Antimicrobial Approach for Pakistan

Abstract

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a significant challenge to global healthcare making the problem of antimicrobial resistance (AMR) even greater. The present study explores how CRISPR-Cas13 armed bacteriophages can be used as an innovative way to fight CRKP. Bacteriophages were designed to contain CRISPR-Cas13, an RNA-targeting system to specifically target the resistance genes of Klebsiella pneumoniae and degrade them. In vitro and in vivo models of the efficacy of this phage CRISPR system showed considerable bacteria growth prevention and recovered antibiotic sensitivity. These findings indicated that phage-CRISPR treatment was permissible to reduce bacteria loads by 95 percent upon time-kill assays and tremendously enhanced survival in Galleria mellonella larvae. Gene expression analysis was used to verify the silencing of carbapenem resistance genes was successful. These results indicate that CRISPR-Cas13-bearing bacteriophages would be useful as a precise therapy in particular in response to multidrug-resistant bacterial infections, which would offer a viable alternative to traditional antibiotic therapy. This will not only help to restore the effectiveness of antibiotics, but will also enhance the availability of a specific antimicrobial treatment that will reduce the chances of resistance being developed.