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The effect of TGF-beta in Apc-mutated mouse intestinal organoids

ABSTRACT: Intestinal crypts isolated from Apcflox/flox; villin-CreERT mice were treated with Tamoxifen to induce the deletion of Apc. Tamoxifen-treated organoids were selected in the absence of Wnt agonists and then treated with TGF-beta.

ORGANISM(S): Mus musculus

PROVIDER: GSE35093 | GEO | 2014/04/18

SECONDARY ACCESSION(S): PRJNA156159

REPOSITORIES: GEO

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The effect of TGF-beta in Apc-mutated mouse intestinal organoids

Project description:Intestinal crypts isolated from Apcflox/flox; villin-CreERT mice were treated with Tamoxifen to induce the deletion of Apc. Tamoxifen-treated organoids were selected in the absence of Wnt agonists and then treated with TGF-beta. Total RNA obtained from Tamoxifen-treated, Apc-deleted intestinal organoids in the absence or presence of 3 ng/ml TGF-beta (18h).

2014-04-18 | E-GEOD-35093 | biostudies-arrayexpress

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Project description:We isolated and selected intestinal adenoma organoids from villin-CreER; Apcflox/flox and villin-CreER; Apcflox/flox; Prox1flox/flox mice and added tamoxifen to induce the deletion of the Apc and Prox1 genes in the intestinal epitheliul ex vivo. Microarray experiments were carried out 7 days after the addition of tamoxifen. Total RNA obtained from villin-CreER; Apcflox/flox and villin-CreER; Apcflox/flox; Prox1flox/flox organoids were compared 7 days after the addition of tamoxifen and 5 days after the selection for Apc-mutant organoids in the absence of the Wnt-agonist R-Spondin1.

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RNAseq comparison of Whole crypt mRNA levels between Control, conditional Notchless KO, Apc KO and Notchless-Apc KO mice 24h or 48h after induction

Project description:Control (Villin-CreERT2tg/0; Apcflox/+; Nleflox/+), ApccKO (Villin-CreERT2tg/0; Apcflox/flox; Nleflox/+) and ApccKO; NlecKO (Villin-CreERT2tg/0; Apcflox/flox; Nleflox/null) mice were subjected to three daily intraperitoneal tamoxifen injections and their intestinal crypts were harvested at 24h (d1) or 48h (d2) post last tamoxifen injection. Total RNA was extracted according to the Trizol-chloroform extraction protocol provided by Invitrogen. RNAseq was performed on these extracted RNA to unravel the molecular mechanisms underlying the ApccKO;NlecKO phenotype.

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Gene expression changes in Apc-mutant mouse intestinal organoids with and without deleting the Prox1 transcription factor

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Gene expression changes in Apc-mutant mouse intestinal organoids 7 and 20 days after the induction of the Apc deletion

Project description:We isolated and selected intestinal adenoma organoids from Lgr5-EGFP-IRES-CreER; Apcflox/flox mice and added tamoxifen to induce the deletion of the Apc gene in the intestinal stem cells. Gene expressions on day7 and day20 after the addition of tamoxifen were compared, representing two stages with different colorectal cancer stem cell content. Total RNA obtained from Lgr5-EGFP-IRES-CreER; Apcflox/flox organoids were compared 7 days and 20 days after the addition of tamoxifen, cultured without the Wnt-agonist R-Spondin1.

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Gene expression changes in Apc-mutant mouse intestinal organoids 7 and 20 days after the induction of the Apc deletion

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Project description:We made intestinal organoid cultures from Villin-CreERT2;Apcflox/flox mice. Organoids were subjected to 4-OH-Tam treatment, whereafter Apc mutant organoids were selected in R-Spondin free medium. DBZ or DMSO treatment was started on day 3. On day 8, organoids were dissociated to obtain single cells and analyzed using the Chromium Single-cell 3'RNA-sequencing system

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Project description:APC is mutated in the majority of colorectal cancers. Inducible deletion of Apc in intestinal epithelial cells in Apcfl//fl; Villin-CreERT2 mice recapitulates this tumor-initiating mutation resulting in expanded intestinal crypts, including stem cells. We used microarrays to analyze BEC gene expression changes during the early stages of intestinal tumorigenesis.

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Expression profile of (inducible) homozygous Apc loss in the mouse intestinal epithelium

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