AspFlex: Molecular Tools to Study Gene Expression and Regulation in .

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 ), which are known to easily develop antibiotic resistances. It is crucial to create a molecular toolkit to investigate its basic biology, such as gene regulation. Despite having been a threat for almost two decades, an efficient and high-throughput plasmid system that can replicate in has not yet been developed. This study adapts an existing toolkit for to meet 's unique requirements and expands it by constructing a plasmid-based CRISPR interference (CRISPRi) system to generate gene knockdowns in .