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Transcriptional response of NGN3 J2 enteroids

ABSTRACT: Transcriptional response of NGN3 J2 enteroids

PROVIDER: PRJNA575547 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10221973_1.fastq.gz	Fastqsanger.gz
	SRR10221973_2.fastq.gz	Fastqsanger.gz
	SRR10221974_1.fastq.gz	Fastqsanger.gz
	SRR10221974_2.fastq.gz	Fastqsanger.gz
	SRR10221975_1.fastq.gz	Fastqsanger.gz

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Transcriptional response of NGN3 J2 enteroids.

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Transcriptional response of NGN3 J2 enteroids

Project description:Enteroendocrine cells are hormonal secreting cells in the gut. However, as they comprise <1% of the total epithelial cell population, studies on their response to stimuli have been limited. Chang-Graham, Danhof, Engevik, and Tomaro-Duchesneau et al (PMID 31029854) developed a model enteroid system to overcome this limitation. By driving expression of NGN3 in human jejunal enteroids, their system allows for analysis of hormonal secretion and transcriptional analysis in response to a stimulus. To further characterize these NGN3 enteroids, we performed RNASeq on these enteroids in the induced or uninduced state.

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Project description:The goal of this experiment is to decipher the mechanism of intestinal endocrine / enteroendocrine subtype specification, that is far from being fully understood. To this end, we used single cell genomics in mouse mini-guts. Enteroendocrine cells are derived from endocrine progenitors expressing the transcription factor Ngn3. We took advantage of the Ngn3+/eYFP mouse model -where endocrine progenitors and their descendants can be isolated and sorted by FACS on the basis of the eYFP fluorescence- and established enteroids or mini-guts from the small intestine. Enteroids were dissociated and eYFP+ single cells were directly sorted in 96 wells of the Precise WTA Single Cell Encoding Plate (BD™ Precise WTA Single Cell Kit, BD Genomics). cDNA and libraries were prepared following the BD protocol. Sequencing (paired-end, 2x100b) was performed in a HiSeq 4000 (Illumina). The bioinformatic analysis allowed to identify 8 different groups of enteroendocrine cells with specific signatures.

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