期刊
SCIENCE
卷 371, 期 6531, 页码 798-+出版社
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.aba5257
关键词
-
资金
- NIH [R01HG009136, 580 R01HG009892, R01DK104908-01]
- Mary Gates Undergraduate Research Scholarship
microSPLiT is a high-throughput single-cell RNA sequencing method that can resolve heterogeneous transcriptional states in Gram-negative and Gram-positive bacteria. Researchers used microSPLiT to process Bacillus subtilis cells and obtained detailed information on changes in metabolism and lifestyle, as well as identified new gene expression states in the bacterial population.
Single-cell RNA sequencing (scRNA-seq) has become an essential tool for characterizing gene expression in eukaryotes, but current methods are incompatible with bacteria. Here, we introduce microSPLiT (microbial split-pool ligation transcriptomics), a high-throughput scRNA-seq method for Gram-negative and Gram-positive bacteria that can resolve heterogeneous transcriptional states. We applied microSPLiT to >25,000 Bacillus subtilis cells sampled at different growth stages, creating an atlas of changes in metabolism and lifestyle. We retrieved detailed gene expression profiles associated with known, but rare, states such as competence and prophage induction and also identified unexpected gene expression states, including the heterogeneous activation of a niche metabolic pathway in a subpopulation of cells. MicroSPLiT paves the way to highthroughput analysis of gene expression in bacterial communities that are otherwise not amenable to single-cell analysis, such as natural microbiota.
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