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Isolation of Mex3a / Lgr5 cell population in mouse small intestine

ABSTRACT: Proximal mouse small intestine from mice bearing the Lgr5 GFP/+ and Mex3a Tom/+ alleles were used to obtain single cell preparations. Cells were selected for GFP expression and different levels of tdTomato were defined. Sorted cells were lysed and processed for transcriptomic analysis

ORGANISM(S): Mus musculus

PROVIDER: GSE90629 | GEO | 2017/06/01

SECONDARY ACCESSION(S): PRJNA355245

REPOSITORIES: GEO

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Transcriptomic profilling of Mex3a+ cells in colon adenomas compared to LGR5

Project description:Transcriptomic data related to 4 different subpopulations found in Mex3a Ki/+ Lgr5 Gfp in APCflfl adenomas in untreated animals. Adenomas where induced with 3%DSS and a single shot of 8mg/kg of Tamoxifen. The populations are refered as Mex3a + Lgr5, Mex3a- Lgr5+ Mex3a+ Lgr5- and Mex3a- Lgr5- according to the flow cytometry profile. Cells were isolated using FACsARIA (BD)

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Mex3a Marks a Slowly Dividing Subpopulation of Lgr5+ Intestinal Stem Cells

Project description:The intestinal epithelium is continuously regenerated by highly proliferative Lgr5+ intestinal stem cells (ISCs). The existence of a population of quiescent ISCs has been suggested yet its identity and features remain controversial. Here we describe that the expression of the RNA-binding protein Mex3a labels a subpopulation of Lgr5+ cells that divide less frequently and contribute to regenerate all intestinal lineages with slow kinetics. Single cell transcriptomic analysis revealed two classes of Lgr5-high cells, one of them defined by the Mex3a-expression program and by low levels of proliferation genes. Lineage tracing experiments show that large fraction of Mex3a+ cell population is continuously recalled into the rapidly dividing self-renewing ISC pool in homeostatic conditions. Chemotherapy and radiation target preferentially rapidly dividing Lgr5+ cells but spare the Mex3a-high/Lgr5+ population, which helps sustain the renewal of the intestinal epithelium during treatment.

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Transcriptomic profiling of small intestinal crypts upon MEX3A deletion

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Transcriptomic profilling of Mex3a+ cells in colon adenomas compared to LGR5

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Distinct populations of embryonic epithelial progenitors generate Lgr5+ intestinal stem cells

Project description:To follow the changes in the transcriptional programs accompanying the specification of the embryonic Lgr5+ cells we sequenced whole transcriptomes of embryonic intestinal epithelium progenitors (at E13.5 and E15.5). EpCAM positive embryonic gut epithelium was isolated from dissected small intestines using fluorescence activated cell sorting (FACS). Lgr5+ progenitors were purified on the basis of GFP fluorescence from Lgr5GFP-Cre-ERT mice (Barker et al. 2007). Double positive embryonic progenitors were isolated by FACS based on GFP and tdTomato fluorescence.

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Mex3a RIP-seq

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Mex3a RIP-seq

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