title: Rapid and Efficient Genome Editing in Staphylococcus aureus by Using an Engineered CRISPR/Cas9 System.
abstract: 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.
authors.affiliation: School of Physical Science and Technology, ShanghaiTech University , Shanghai 201210, China.
authors.forename: Weizhong
authors.initials: W
authors.lastname: Chen
authors.affiliation: School of Physical Science and Technology, ShanghaiTech University , Shanghai 201210, China.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Microbiology and Immunology, Indiana University, School of Medicine-Northwest , Gary, Indiana 46408, United States.
authors.forename: Won-Sik
authors.initials: WS
authors.lastname: Yeo
authors.affiliation: Department of Microbiology and Immunology, Indiana University, School of Medicine-Northwest , Gary, Indiana 46408, United States.
authors.forename: Taeok
authors.initials: T
authors.lastname: Bae
authors.affiliation: School of Physical Science and Technology, ShanghaiTech University , Shanghai 201210, China.
authors.forename: Quanjiang
authors.identifier: 0000-0002-2321-8462
authors.initials: Q
authors.lastname: Ji
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D000072669:Gene Editing; D005818:Genetic Engineering; D009154:Mutation; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics