AspFlex: Molecular Tools to Study Gene Expression and Regulation in Acinetobacter baumannii

Acinetobacter baumannii is a Gram-negativenosocomial opportunistic pathogen frequently found in hospital settings,causing high incidence of in-hospital infections. It belongs to theESKAPE group of pathogens (the "A" stands for A. baumannii), which are known to easily developantibiotic resistances. It is crucial to create a molecular toolkitto investigate its basic biology, such as gene regulation. Despite A. baumannii having been a threat for almost twodecades, an efficient and high-throughput plasmid system that canreplicate in A. baumannii has not yetbeen developed. This study adapts an existing toolkit for Escherichia coli to meet A. baumannii's unique requirements and expands it by constructing a plasmid-basedCRISPR interference (CRISPRi) system to generate gene knockdowns in A. baumannii.