Single-cell RNA sequencing using split-pool barcoding reveals transcriptional heterogeneity in with implications for periodontal pathogenesis.

Background: is a keystone pathogen in periodontitis, associated with dysbiosis and chronic inflammation. While its virulence mechanisms are well characterized, its transcriptional heterogeneity at the single-cell level remains unexplored.

Materials And Methods: We applied split-pool barcoding-based single-cell RNA sequencing to profile gene expression in 1,942 individual W83 cells cultured under anaerobic conditions. Clustering and differential expression analyses were conducted to identify distinct transcriptional subpopulations.

Results: We identified six transcriptionally distinct clusters, with the two largest accounting for 72.7% of the population. Minor clusters exhibited signatures related to stress responses, metabolism, membrane transport, and DNA regulation. Sub-clustering of major populations revealed rare subgroups, including one enriched for genes involved in iron acquisition, proteolysis, and transport.

Conclusions: This study presents the first single-cell transcriptomic map of , revealing rare but functionally significant subpopulations. Such diversity may support bacterial adaptability, virulence, and immune evasion, informing future strategies for targeted periodontal therapy.