Rapid and Efficient Genome Editing in Staphylococcus aureus by Using an Engineered CRISPR/Cas9 System.
Weizhong ChenYifei ZhangWon-Sik YeoTaeok BaeQuanjiang JiPublished in: Journal of the American Chemical Society (2017)
Staphylococcus aureus, a major human pathogen, has been the cause of serious infectious diseases with a high mortality rate. Although genetics is a key means to study S. aureus physiology, such as drug resistance and pathogenesis, genetic manipulation in S. aureus is always time-consuming and labor-intensive. Here we report a CRISPR/Cas9 system (pCasSA) for rapid and efficient genome editing, including gene deletion, insertion, and single-base substitution mutation in S. aureus. The designed pCasSA system is amenable to the assembly of spacers and repair arms by Golden Gate assembly and Gibson assembly, respectively, enabling rapid construction of the plasmids for editing. We further engineered the pCasSA system to be an efficient transcription inhibition system for gene knockdown and possible genome-wide screening. The development of the CRISPR/Cas9-mediated genome editing and transcription inhibition tools will dramatically accelerate drug-target exploration and drug development.
Keyphrases
- crispr cas
- genome editing
- genome wide
- staphylococcus aureus
- infectious diseases
- copy number
- dna methylation
- loop mediated isothermal amplification
- endothelial cells
- escherichia coli
- transcription factor
- biofilm formation
- risk factors
- cardiovascular events
- type diabetes
- methicillin resistant staphylococcus aureus
- coronary artery disease
- pseudomonas aeruginosa
- sensitive detection
- high density