Single-cell RNA sequencing of Porphyromonas gingivalis reveals heterogeneity and rare cellular states in laboratory medium.

Porphyromonas gingivalis is a Gram-negative anaerobic bacterium linked to chronic periodontitis and systemic diseases. Understanding its cellular heterogeneity is crucial for elucidating its adaptive mechanisms and virulence regulation. This study employs single-cell RNA sequencing using the split-pool barcoding method to capture transcriptomic variability within P. gingivalis populations. The results provide insights into gene expression diversity under laboratory growth conditions, revealing potential subpopulations that contribute to bacterial survival and pathogenicity. Overall design: Single-cell RNA sequencing (scRNA-seq) was performed on P. gingivalis W83 using PETRI-seq, a split-pool barcoding approach optimized for bacterial transcriptomics. The bacterial culture was grown to mid-log phase, followed by fixation, permeabilization, and RNA processing. Barcoded cDNA libraries were prepared, amplified, and sequenced using the Illumina NextSeq platform. Data analysis focused on identifying distinct transcriptional states and heterogeneity within the bacterial population.