Integrated single-cell transcriptomic and network analysis identifies key regulators of Acinetobacter baumannii biofilm integrity

Biofilm growth curves of Acinetobacter baumannii were determined using the crystal violet staining method. Gene expression differences among different cell clusters were compared based on single-cell RNA sequencing data and validated by RT-qPCR. Functional validation of the target gene was further performed by comparing biofilm formation between wild-type (WT) and gene-knockout (KO) strains. Compared to the WT strain, the KO strain exhibited significantly reduced biofilm biomass and permeability, confirming the gene's role as a key regulator in biofilm formation. Targeting this gene may disrupt biofilm-associated drug resistance, providing a novel strategy for treating persistent infections.采用结晶紫染色法测定鲍曼不动杆菌（Acinetobacter baumannii）的生物膜生长曲线。基于单细胞RNA测序（single-cell RNA sequencing）数据，比较不同细胞簇间的基因表达差异，并通过实时定量逆转录聚合酶链反应（RT-qPCR）进行验证。进一步通过比较野生型（WT）与基因敲除（KO）菌株的生物膜形成能力，完成靶基因的功能验证。与野生型菌株相比，基因敲除菌株的生物膜生物量与通透性均显著降低，证实该基因是生物膜形成过程中的关键调控因子。靶向该基因可干扰生物膜相关耐药性，为持续性感染的治疗提供全新策略。