ChIPseq of enterocytes of the jejunum from transgene C57BL/6 mice harboring a CKIαΔgut knock-out
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ABSTRACT: ChIPseq of enterocytes of the jejunum from transgene C57BL/6 mice to assess the effects of a CKIαΔgut knock-out in the jejunum and a CKIαΔgut/p53 double knock-out. In addition, enterocytes expressing a mutant p53 (R172H) in combination with CKIαΔgut knockout were analyzed.
Project description:RNAseq of enterocytes of the jejunum and Ileum from transgene C57BL/6 mice to assess the effects of a CKIα knock-out in combination with expression of a p53 mutant protein (R172H ) compared to a double knock-out of CKIα and p53. In addition, a group of mice exhibiting a CKIα knock-out and mutant p53 (R172H) was treated with gallic acid and compared to untreated mice (Jejunum only).
Project description:The unmodified DNA fraction of jejunal enterocytes and the jejunum lacking enterocytes of humans and jejunal enterocytes of mice was interrogated on Affymetrix tiling arrays The mTAG technique was used to enrich the unmodified DNA fraction in a total of 56 humans and 26 mice
Project description:To elucidate the effect of senescence in mouse colorectal tumor, the transcriptome of enterocytes from CKI alpha deletion mutants (CKI alpha KO), CKI alpha/p53 double deletion mutants (CKI alpha/p53 DKO) and CKI alpha heterozygous mice (CKI alpha Het) were determined by RNAseq.
Project description:This experiment was performed to determine which gene promoters mutant p53 binds and transcriptionally regulates in order to understand how mutant p53 accomplishes its gain of function phenotype.
Project description:The mouse R178E (EE) mutation of the Trp53 is a p53 mutant protein with native conformation that lacks the ability to form tetramers and thus constitutes a mutant form of p53 that lacks DNA binding cooperativity. Here we want to assess DNA binding ability of the EE mutant in MEFs under untreated conditions and following p53 stabilization with the Mdm2 inhibitor nutlin-3a and compare it to p53 KO and WT mice.
Project description:Intestinal epithelial spheroids were isolated from mouse jejunum as described in our previous publication (Miyoshi and Stappenbeck. In vitro expansion and genetic modification of gastrointestinal stem cells in spheroid culture. Nature Protocols. 2013. PMID: 24232249) and then grown in 50% L-WRN conditioned medium (for stem cells) or differentiation medium containing dmPGE2 (for wound-associated epithelial cells) or differentiation medium containing EP4 inhibitor (for enterocytes). Transcription profiling was performed to gain insight into the characteristics distinguishing these cell types.
Project description:SV40 large T antigen (TAg) contributes to cell transformation, in part, by targeting two well characterized tumor suppressors, pRb and p53. TAg expression affects the transcriptional circuits controlled by Rb and by p53. We have performed a microarray analysis to examine the global change in gene expression induced by wild-type TAg and TAg-mutants, in an effort to link changes in gene expression to specific transforming functions. For this analysis we have used enterocytes from the mouse small intestine expressing TAg. Expression of TAg in the mouse intestine results in hyperplasia and dysplasia. Our analysis indicates that practically all gene expression regulated by TAg in enterocytes is dependent upon its binding and inactivation of the Rb-family proteins. Experiment Overall Design: Laser capture microdissection (LCM) was used to isolate villus enterocytes from three independent non-transgenic mice, four wild-type TAg transgenic mice, five N136 transgenic mice, three D44N transgenic mice and three 3213 transgenic mice. Total RNA was extracted from the enterocytes with the Pico Pure kit and amplified twice with the Ribo Amp kit (Arcturus). Amplified RNA was processed by the Genomics and Proteomics Core Laboratories at the University of Pittsburgh and hybridized to the Affymetrix GeneChip Mouse Genome 430 2.0 array. MAS 5.0 was used to obtain the present/absent calls and CEL files were normalized by RMA to obtain log2 expression values.
Project description:The unmodified DNA fraction of jejunal enterocytes and the jejunum lacking enterocytes of humans and jejunal enterocytes of mice was interrogated on Affymetrix tiling arrays
Project description:This SuperSeries is composed of the following subset Series: GSE41541: Expression data from mouse proximal intestinal epithelial Lgr5(hi) stem cells and differentiated villus cells (enterocytes from Atoh1 conditional knockout) GSE41542: H3K79me2 ChIP-seq in mouse proximal intestinal Lgr5(hi) stem cells and villus cells GSE41710: Global gene expression analysis of Dot1l-deficient and control intestinal villus cells in mouse Refer to individual Series