AspFlex: molecular tools to study gene expression and regulation in Acinetobacter baumannii

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. baumanniis unique requirements and expands it by constructing a mobile CRISPR interference (CRISPRi) system that can produce gene knockdowns in A. baumannii.