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Single-cell sequencing of human intestnal organoids with and without enteroendocrine cells (EECs)

ABSTRACT: Single-cell sequencing of human intestnal organoids with and without enteroendocrine cells (EECs)

PROVIDER: PRJNA887399 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21815034.fastq.gz	Fastqsanger.gz
	SRR21815035.fastq.gz	Fastqsanger.gz
	SRR21815036.fastq.gz	Fastqsanger.gz
	SRR21815037.fastq.gz	Fastqsanger.gz
	SRR21815038.fastq.gz	Fastqsanger.gz

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Single-cell sequencing of human intestnal organoids with and without enteroendocrine cells (EECs)

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Project description:Homeostatic regulation of the intestinal enteroendocrine lineage hierarchy is a poorly understood process. We resolved transcriptional changes during enteroendocrine differentiation in real-time at single-cell level using a novel knock-in allele of Neurog3, the master regulator gene briefly expressed at the onset of enteroendocrine specification. A bi-fluorescent reporter, Neurog3Chrono, measures time from the onset of enteroendocrine differentiation and enables precise positioning of single-cell transcriptomes along an absolute time axis. This approach yielded a definitive description of the enteroendocrine hierarchy and its sub-lineages, uncovered differential kinetics between sub-lineages, and revealed time-dependent hormonal plasticity in enterochromaffin and L-cells. The time-resolved map of transcriptional changes predicted multiple novel molecular regulators. Nine of these were validated by conditional knockout in mice or CRISPR-modification in intestinal organoids. Six novel candidate regulators (Sox4, Rfx6, Tox3, Myt1, Runx1t1 and Zcchc12) yielded specific enteroendocrine phenotypes. Our time-resolved single cell transcriptional map presents a rich resource to unravel enteroendocrine differentiation.

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