SLAM-ITseq: sequencing cell type-specific transcriptomes without cell sorting

Cell type-specific transcriptome analysis is an essential tool for understanding biological processes in which diverse types of cells are involved. Although cell isolation methods such as fluorescence-activated cell sorting (FACS) in combination with transcriptome analysis have widely been used so f...

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Veröffentlicht in:Development (Cambridge) 2018-07, Vol.145 (13), p.dev164640-dev164640
Hauptverfasser: Matsushima, Wayo, Herzog, Veronika A, Neumann, Tobias, Gapp, Katharina, Zuber, Johannes, Ameres, Stefan L, Miska, Eric A
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Sprache:eng
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Zusammenfassung:Cell type-specific transcriptome analysis is an essential tool for understanding biological processes in which diverse types of cells are involved. Although cell isolation methods such as fluorescence-activated cell sorting (FACS) in combination with transcriptome analysis have widely been used so far, their time-consuming and harsh procedures limit their applications. Here, we report a novel metabolic RNA sequencing method, SLAM-ITseq, which metabolically labels RNA with 4-thiouracil in a specific cell type followed by detection through an RNA-seq-based method that specifically distinguishes the thiolated uridine by base conversion. This method has successfully identified the cell type-specific transcriptome in three different tissues: endothelial cells in brain, epithelial cells in intestine and adipocytes in white adipose tissue. As this method does not require isolation of cells or RNA prior to the transcriptomic analysis, SLAM-ITseq provides an easy yet accurate snapshot of the transcriptional state .
ISSN:0950-1991
1477-9129
DOI:10.1242/dev.164640