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Single cell RNA-seq of Sox10-Cre::GFP+ population

ABSTRACT: Single cell RNA-seq of Sox10-Cre::GFP+ population

PROVIDER: PRJNA686369 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13272124.fastq.gz	Fastqsanger.gz
	SRR13272125.fastq.gz	Fastqsanger.gz
	SRR13272126.fastq.gz	Fastqsanger.gz
	SRR13272127.fastq.gz	Fastqsanger.gz

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Single cell RNA-seq of Sox10-Cre::GFP+ population

Project description:We performed single-cell RNA-seq on GFP+ sorted population from the brain from the Sox10-Cre::GFP transgenic mouse at postnatal day P21

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Determination of Sox10-regulated genes in the oligodendroglial precursor cell line OLN93

Project description:To determine Sox10-regulated genes in the oligodendroglial precursor cell line OLN93, CRISPR/Cas9-mediated genome editing was performed to inactivate Sox10. For CRISPR/Cas9-mediated Sox10 inactivation, two guide sequences (5’-GCTGCGAGCCAGCCCAGGGCC-3’ for clone A and 5’-GTTGCCCAATTCGCCG GGCCC-3’ for clone B) were separately cloned into pX330. Oln93 cells were co-transfected with pEGFP-N1 and the pX330 plasmids that express the respective guide RNA and SpCas9. Transfected GFP-positive cells were enriched by fluorescence activated cell-sorting and seeded at single cell density in 96-well plates. Homogeneously GFP-expressing clones were analysed for loss of Sox10 protein by immunocytochemistry. Clones without Sox10 protein were obtained with both guide RNAs. Sox10 inactivation was confirmed by Sanger sequencing of the Sox10 gene. RNA was isolated from one clone A and one clone B as well as two independent RNA samples of wildtype OLN93 cells and submitted to RNA-seq.

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Project description:We have recently demonstrated that Sox10-expressing (Sox10+) cells give rise to mainly type-III neuronal taste bud cells that are responsible for sour and salt taste. The two tissue compartments containing Sox10+ cells in the surrounding of taste buds include the connective tissue core of taste papillae and von Ebner’s glands (vEGs) that are connected to the trench of circumvallate and foliate papillae. Here we present data to support that it is the vEGs, not connective tissue core, that serve as the niche of Sox10+ taste bud progenitors. In this study, we performed single cell RNA-sequencing of the epithelium of Sox10-Cre/tdT mouse circumvallate/vEG complex and used inducible Cre mouse models to map the cell lineages of vEGs and/or connective tissue (including stromal and Schwann cells). Transcriptomic analysis indicated that Sox10 expression was enriched in the cell clusters of vEG ducts that contained abundant proliferating cells, while Sox10-Cre/tdT expression was enriched in type-III taste bud cells and vEG ductal cells. In vivo lineage mapping showed that the traced cells were distributed in circumvallate taste buds concurrently with those in the vEGs, but not in the connective tissue. Moreover, multiple genes encoding pathogen receptors were enriched in the vEG ducts hosting Sox10+ cells. If this is also true in humans, our data indicates that vEG duct is a source of Sox10+ taste bud progenitors and susceptible to pathogen infections.

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