Rapid and Efficient Genome Editing in Staphylococcus aureus by Using an Engineered CRISPR/Cas9 System

Weizhong Chen, Yifei Zhang, Won Sik Yeo, Taeok Bae, Quanjiang Ji

Research output: Contribution to journalArticle

32 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)3790-3795
Number of pages6
JournalJournal of the American Chemical Society
Volume139
Issue number10
DOIs
StatePublished - Mar 15 2017

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Staphylococcus aureus
Genes
Transcription
Gene Knockdown Techniques
Insertional Mutagenesis
Pharmaceutical Preparations
Gene Deletion
Drug Resistance
Communicable Diseases
Physiology
Pathogens
Plasmids
Genome
Screening
Repair
Substitution reactions
Mutation
Mortality
Gene Editing

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Rapid and Efficient Genome Editing in Staphylococcus aureus by Using an Engineered CRISPR/Cas9 System. / Chen, Weizhong; Zhang, Yifei; Yeo, Won Sik; Bae, Taeok; Ji, Quanjiang.

In: Journal of the American Chemical Society, Vol. 139, No. 10, 15.03.2017, p. 3790-3795.

Research output: Contribution to journalArticle

Chen, Weizhong ; Zhang, Yifei ; Yeo, Won Sik ; Bae, Taeok ; Ji, Quanjiang. / Rapid and Efficient Genome Editing in Staphylococcus aureus by Using an Engineered CRISPR/Cas9 System. In: Journal of the American Chemical Society. 2017 ; Vol. 139, No. 10. pp. 3790-3795.
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