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Single-cell massively-parallel multiplexed microbial sequencing (M3-seq) identifies rare bacterial populations and profiles phage infection

Author(s): Wang, Bruce; Lin, Aaron E; Yuan, Jiayi; Koch, Matthias D; Adamson, Britt; et al

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Abstract: Bacterial populations are highly adaptive. They can respond to stress and survive in shifting environments. How the behaviours of individual bacteria vary during stress, however, is poorly understood. To identify and characterize rare bacterial subpopulations, technologies for single-cell transcriptional profiling have been developed. Existing approaches show some degree of limitation, for example, in terms of number of cells or transcripts that can be profiled. Due in part to these limitations, few conditions have been studied with these tools. Here we develop massively-parallel, multiplexed, microbial sequencing (M3-seq)—a single-cell RNA-sequencing platform for bacteria that pairs combinatorial cell indexing with post hoc rRNA depletion. We show that M3-seq can profile bacterial cells from different species under a range of conditions in single experiments. We then apply M3-seq to hundreds of thousands of cells, revealing rare populations and insights into bet-hedging associated with stress responses and characterizing phage infection.
Publication Date: 31-Aug-2023
Citation: Wang, Bruce, Lin, Aaron E, Yuan, Jiayi, Koch, Matthias D, Adamson, Britt, Wingreen, Ned S, Gitai, Zemer. (2022). Massively-parallel Microbial mRNA Sequencing (M3-Seq) reveals heterogeneous behaviors in bacteria at single-cell resolution. Cold Spring Harbor Laboratory, 10.1101/2022.09.21.508688
Type of Material: Journal Article
Journal/Proceeding Title: Nature Microbiology
Version: Final published version. This is an open access article.

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