Abstract
Acinetobacter baumannii is a gram negative nosocomial opportunistic pathogen frequently found in hospital settings, causing high incidence of in-hospital infections. It belongs to the ESKAPE group of pathogens (the “A” stands for A. baumannii) that is known to easily develop antibiotic resistances. It is crucial to create a molecular toolkit to investigate its basic biology, such as gene regulation. Despite A. baumannii being a threat for almost two decades, an efficient and high-throughput plasmid system that can replicate in A. baumannii has not yet been developed. This study adapts an existing toolkit for Escherichia coli to meet A. baumannii’s unique requirements and expands it by constructing a mobile CRISPR interference (CRISPRi) system that can produce gene knockdowns in A. baumannii.
Competing Interest Statement
The authors have declared no competing interest.
Abbreviations
- ESKAPE
- Enterococcus faecium; Staphylococcus aureus; Klebsiella pneumoniae; Acinetobacter baumannii; Pseudomonas aeruginosa; Enterobacter spp 6 organisms of critical importance in inhospital infections because of their abilities to gain antibiotic resistances.
- MoClo
- Modular Cloning. Plasmids sets specifically created for Golden Gate cloning.
- CRISPRi
- Clustered regularly interspaced short palindromic repeats inhibition. An expression inhibition system based on a deactivated Cas9 protein physically blocking transcription of a genomic region through an sgRNA binding in that region.
- egfp
- enhanced green fluorescent protein.
- ATc
- Anhydrous Tetracycline
- X-Ga1
- 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside. An organic compound used for blue-white screening.
