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RNA-seq of Paneth cells

ABSTRACT: RNA-seq of Paneth cells

PROVIDER: PRJNA736960 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14788304.fastq.gz	Fastqsanger.gz
	SRR14788305.fastq.gz	Fastqsanger.gz
	SRR14788306.fastq.gz	Fastqsanger.gz
	SRR14788307.fastq.gz	Fastqsanger.gz
	SRR14788308.fastq.gz	Fastqsanger.gz

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RNA-seq of Paneth cells

Project description:The impact of removal of Mll1 in intestinal Paneth cells expressing an constitutively active form of beta-catenin (beta-cateninGOF) was analysed in 4 pairs of sorted Paneth cells of Lgr5-CreERT2; beta-cateninGOF;Mll1+/- (control) and Lgr5-CreERT2; beta-cateninGOF;Mll1-/- (knockout) mice at 10 days after tamoxifen-induced mutagenesis. Using 75-base-pair reads, around 30 million reads per sample with comparable unique mapped reads (52-93%) were obtained. To analyze differentially expressed genes, we applied DESeq2 analysis to the RNA-seq dataset. Differentially expressed genes in beta-cateninGOF; Mll1-/- versus beta-cateninGOF; Mll1+/- Paneth cells showed both up- and downregulation of genes at a false discovery rate (FDR) of 10%. This included a global increase in the expression of goblet cell-specific genes. Downregulated genes included specific markers of Paneth cells, indicating that ablation of Mll1 in Paneth cells shifted their identity towards a mixed Paneth-goblet cell fate.

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RNA-seq of sorted intestinal stem and Paneth cells

Project description:RNA-seq of sorted intestinal stem and Paneth cells

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RNA-seq of sorted intestinal stem and Paneth cells

Project description:The impact of Mll1 removal on the intestinal stem cells and its direct effect on neighbouring Paneth cells was evaluated in sorted intestinal stem and Paneth cells from Mll1FC/+; Lgr5-eGFP-CreERT2/+ (control) and Mll1FC/FC; Lgr5-eGFP-CreERT2/+ (knockout) mice, 4 and 10 days after tamoxifen-induced mutagenesis. Using 75-base-pair reads, 30 million reads per sample with comparable unique mapped reads for stem (70-77%) and Paneth (60-76%) cells were obtained. To analyze differentially expressed genes, we applied DESeq2 analysis to the RNA-seq dataset. Analysis by DAVID and GSEA at a false discovery rate (FDR) of 5% was conducted.The stem cell transcriptome revealed that Mll1 knockout stem cells exhibited a decreased expression of several transcription factors and stem cell genes. Additionally, Mll1 ablation in stem cells had an impact on Paneth cells. Downregulation of Paneth cell specific markers indicated a loss of Paneth cell identity.

2021-11-10 | GSE157285 | GEO

Single cell RNA sequencing analysis of germ free (GF) mouse Paneth cells and microbiome-transferred exGF mouse Paneth cells.

Project description:The single cell level Paneth cell transcriptome under gnotobiotic condition has not been resolved. How gut microbiome modulates Paneth cell transcriptome at single cell level in germ-free mice was not known. We used a flow cytometry method to isolate highly pure ileal Paneth cells from germ free (GF) Paneth cell reporter mice (Lyz1-3'UTR-IRES-CreER) and from exGF Paneth cell reporter mice that were transplanted with wild type C57B/l6 mouse gut microbiota (exGF+B6M). These isolated Paneth cells were subjected to single cell RNA sequencing by 10xGenomics.

2023-09-01 | GSE237328 | GEO

RNA-seq of mouse Paneth and Lgr5 expressing intestinal stem cells (ISCs)

Project description:RNA-seq of mouse Paneth and Lgr5 expressing intestinal stem cells (ISCs)

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Mouse duodenal and ileal Paneth cell single cell RNA sequencing

Project description:An indepth analysis of Paneth cell transcriptome at single cell level has not been available. Existing intestinal epithelial cell scRNA dataset contain many cell types, where Paneth cells represent a small portion. We used a flow cytometry based approach to enrich and isolate relatively pure Panethc cells from a newly developed Paneth cell reporter mouse line (Lyz1-3'UTR-IRES-CreER; Rosa26R-tdTomato). Single cell RNA sequencing was performed on purified duodenal and ileal Paneth cells of mice housed under specific pathogen free condition.

2023-09-01 | GSE237327 | GEO

Paneth cell scRNA-seq analysis of un-infected and Salmonella Typhimurium infected mice

Project description:Paneth cells consist of heterogeous phenotypes. How Paneth cells respond to infection by invasive pathogenic bacteria at single cell level remains uncharacterized. We used a flow cytometry based method to isolate Paneth cells from uninfected and Salmonella-infected Paneth cell reporter mice (Lyz1-3'UTR-IRES-CreER; Rosa26R-tdTomato). These highly pure Paneth cell populations were used for single cell RNA sequencing analysis.

2023-09-01 | GSE237326 | GEO

RNA-seq of mouse Paneth and Lgr5 expressing intestinal stem cells (ISCs)

Project description:The experiment shows the aging related expression changes in Paneth and Lgr5Hi cells between young and old mice. These expression changes are related to the role of stem cell niche in aging and regeneration of intestinal epithelium.

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Proteomic profiling of mouse Paneth cells microdissected from small intestinal tissue

Project description:Paneth cells are antimicrobial peptide-secreting cells located at the base of the crypts of the small intestine. The proteome of Paneth cells is not well defined because of their co-existence with stem cells making it difficult to culture Panth cells alone in vitro. Using a simplied toluidine blue O method for staining mouse intestinal tissue, laser capture microdissection (LCM) to isolate cells from the crypt region and surfactant assisted one pot protein digestion, we identified more than 1,300 proteins from crypts equivalent to 18,000 cells. Compared with the proteomes of villi and smooth muscle regions, the crypt proteome is highly enriched in defensins, lysozymes and other antimicrobial peptides that are characteristic of Paneth cells. The sensitivity of the LCM-based proteomics approach was also assessed using a smaller number of cell equivalent tissues, a comparable proteomic coverage can be achieved with 3,600 cells. This work is the first proteomics study of intestinal tissue enriched with Paneth cells. The simplied workflow enables profiling of Paneth cell associated pathological changes at the proteome level directly from frozen intestinal tissue. It may also be useful for proteomics studies of other spatially resolved cell types from other tissues.

2022-10-12 | PXD034143 | Pride

RNA-seq of mouse ileum to investigate the role of IL-17 receptor signaling in Paneth cells in their biology and intestinal physiology

Project description:IL-17 receptor plays a major role in the antibacterial defense and regulation of host-microbiota interaction. However its impact on Paneth cells, cell-type specific for production of antibacterial peptides, is still not clear. Here, we used genetic depletion of IL-17 receptor specific for Paneth cell population (Defa6-iCre x Il17ra-flox) and performed the RNA sequencing on FACS-sorted Paneth cells and also complete ileum tissue, comparing Cre+ and Cre- littermates.

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