137
DNA. Guide RNA can be redesigned in such a way that it recognizes the gene of
interest (antimicrobial resistance causing genes) and facilitate the Cas9 nuclease
complex to reach the target. In an experiment, researchers transformed the plasmid
coding for CRISPR-Cas9 guide RNA into E. coli and Staphylococcus which resulted
in the cessation of bacterial growth possessing antimicrobial resistance causing
gene in the presence of antibiotics. The results advocate the presence of transformed
plasmid facilitating degradation of antimicrobial resistance gene (Jinek et al. 2012;
Citorik et al. 2014; Shabbir et al. 2019). Target (antibiotic resistant gene) oriented
Cas9 assembly has better toxicity towards the bacterial cell. CRISPR-Cas9 nuclease
assembly can be used to target the antimicrobial resistance genes and help in resensitization of bacteria towards the antimicrobial agent (Bikard et al. 2014; Diep
et al.J 2006). In a recent study, re-sensitization of Shewanella algae to carbapenem
using CRISPR-Cas9 gene edition was reported (Wu et al. 2019).
Table 6.2 Limitations of commonly used nucleases for gene editing
Type of Nuclease
Limitations
References
Meganuclease
Low target specificity
Hsu et al. (2014) and Shabbir
et al. (2019)
Zinc finger
nucleases
It is arduous to design
Narrow range of target
Shabbir et al. (2019)
Trans activator like
effector
Uncomplicated design and highly
specific to target but it in large in size
Miller et al. (2011)
Difficultly in insertion inside the cell
Strong and Musunuru (2016) and
Shabbir et al. (2019)
Fig. 6.2 CRISPR-Cas9 to overcome antimicrobial resistance in bacteria. The CRISPR-Cas9
system can be delivered in bacteria either using nanoparticles or bacteriophages. The redesigned
guide RNA facilitates the CRISPR-Cas system to the gene responsible for antimicrobial resistance.
The CRISPR nuclease complex disrupts the gene and results in re-sensitization of bacteria towards
antimicrobial agent
6 Role of Gene Editing Tool CRISPR-Cas in the Management of Antimicrobial…
DNA. Guide RNA can be redesigned in such a way that it recognizes the gene of
interest (antimicrobial resistance causing genes) and facilitate the Cas9 nuclease
complex to reach the target. In an experiment, researchers transformed the plasmid
coding for CRISPR-Cas9 guide RNA into E. coli and Staphylococcus which resulted
in the cessation of bacterial growth possessing antimicrobial resistance causing
gene in the presence of antibiotics. The results advocate the presence of transformed
plasmid facilitating degradation of antimicrobial resistance gene (Jinek et al. 2012;
Citorik et al. 2014; Shabbir et al. 2019). Target (antibiotic resistant gene) oriented
Cas9 assembly has better toxicity towards the bacterial cell. CRISPR-Cas9 nuclease
assembly can be used to target the antimicrobial resistance genes and help in resensitization of bacteria towards the antimicrobial agent (Bikard et al. 2014; Diep
et al.J 2006). In a recent study, re-sensitization of Shewanella algae to carbapenem
using CRISPR-Cas9 gene edition was reported (Wu et al. 2019).
Table 6.2 Limitations of commonly used nucleases for gene editing
Type of Nuclease
Limitations
References
Meganuclease
Low target specificity
Hsu et al. (2014) and Shabbir
et al. (2019)
Zinc finger
nucleases
It is arduous to design
Narrow range of target
Shabbir et al. (2019)
Trans activator like
effector
Uncomplicated design and highly
specific to target but it in large in size
Miller et al. (2011)
Difficultly in insertion inside the cell
Strong and Musunuru (2016) and
Shabbir et al. (2019)
Fig. 6.2 CRISPR-Cas9 to overcome antimicrobial resistance in bacteria. The CRISPR-Cas9
system can be delivered in bacteria either using nanoparticles or bacteriophages. The redesigned
guide RNA facilitates the CRISPR-Cas system to the gene responsible for antimicrobial resistance.
The CRISPR nuclease complex disrupts the gene and results in re-sensitization of bacteria towards
antimicrobial agent
6 Role of Gene Editing Tool CRISPR-Cas in the Management of Antimicrobial…