Microarray profiling to analyze the effect of Snai1 loss in mouse intestinal epithelium

Abstract

Epithelial stem cells from a variety of tissues have been shown to express genes linked to mesenchymal cell states. The Snail family of transcriptional factors has long been regarded as a marker of mesenchymal cells, however recent studies have indicated an involvement in regulation of epithelial stem cell populations. Snai1 is expressed in the stem cell population found at the base of the mouse small intestinal crypt that is responsible for generating all differentiated cell types of the intestinal epithelium. We utilized an inducible Cre recombinase approach in the intestinal epithelium combined with a conditional floxed Snai1 allele to induce knockout of gene function in the stem cell population. Loss of Snai1 resulted in loss of crypt base columnar cells and a failure to induce a proliferative response following radiation damage. We induced Snai1 loss in cultured organoids that had been derived from epithelial cells and compared gene expression to organoids with functional Snai1. Here we describe in detail the methods for generation of knockout organoids and analysis of microarray data that has been deposited in Gene Expression Omnibus (GEO):GSE65005.

Keywords: Intestine; SerinC3; Snai1; Stem cells.

Fig. 1

Droplet digital PCR of SerinC3 (normalized to HPRT) expression in VillinCreERT2 Snai1^fl/fl intestinal crypts, 5 days after tamoxifen treatment, compared to control crypts, VillinCreERT2 Snai1^+/+. p = 0.004, Student's T-test.