An Organoid-based CRISPR-Cas9 Screen for Regulators of Intestinal Epithelial Maturation and Cell Fate

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2023 Jul 12

PMID 37436979

Authors

Stine L Hansen

Hjalte L Larsen

Laura M Pikkupeura

Grzegorz Maciag

Jordi Guiu

Iris Muller

Ditte L Clement

Christina Mueller

Jens Vilstrup Johansen

Kristian Helin

Mads Lerdrup

Kim B Jensen

Affiliations

Soon will be listed here.

Abstract

Generation of functionally mature organs requires exquisite control of transcriptional programs governing cell state transitions during development. Despite advances in understanding the behavior of adult intestinal stem cells and their progeny, the transcriptional regulators that control the emergence of the mature intestinal phenotype remain largely unknown. Using mouse fetal and adult small intestinal organoids, we uncover transcriptional differences between the fetal and adult state and identify rare adult-like cells present in fetal organoids. This suggests that fetal organoids have an inherent potential to mature, which is locked by a regulatory program. By implementing a CRISPR-Cas9 screen targeting transcriptional regulators expressed in fetal organoids, we establish and as important factors safeguarding the immature progenitor state. Our approach demonstrates the utility of organoid models in the identification of factors regulating cell fate and state transitions during tissue maturation and reveals that SMARCA4 and SMARCC1 prevent precocious differentiation during intestinal development.

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