Project description:We used micro-dissection with FACS sorting techniques to isolate renal vesicle single cell types from post natal (P4) kidneys. A subset of these single cell populations is analysed individually via Fluidigm single cell analysis. This analysis will determine the transcriptional profile of each cell type, identify compartment specific transcripts, compartment specific transcript isoforms and cell-type specific long-noncoding RNAs. In addition the unbiased nature of RNA-SEQ will potentially identify novel transcripts that have not been annotated in the database. Kidneys are harvested from Tg(Crym-EGFP)GF82Gsat mice. Single cells are extracted from P4 renal vesicles using micro-dissection with FACS sorting techniques. A subset of these cells is analyzed individually via Fluidigm single cell analysis. The long term goal is to generate a transcriptional atlas of the developing kidney.
Project description:Gene expression profiles of cap mesenchyme and renal vesicle isolated between P0-P4 from Crym-EGFP neonatal transgenic mice using FACS. (GUDMAP Series ID: 28)
Project description:We used micro-dissection with FACS sorting techniques to isolate single cells from the metanephric mesenchyme of the E11.5 developing kidney. A subset of these single cell populations is analysed individually via Fluidigm single cell analysis. This analysis will determine the transcriptional profile of each cell type, identify compartment specific transcripts, compartment specific transcript isoforms and cell-type specific long-noncoding RNAs. In addition the unbiased nature of RNA-SEQ will potentially identify novel transcripts that have not been annotated in the database. Kidneys are harvested from Tg(Crym-EGFP)GF82Gsat mice. Single cells are extracted from E11.5 metanephric mesenchyme using manual micro-dissection techniques. A subset of these cells is analyzed individually via Fluidigm single cell analysis. The long term goal is to generate a transcriptional atlas of the developing kidney.
Project description:To identify the potential interaction partners of PRSS37, three testis lysates of Prss37E/+ mice (PRSS37-EGFP knock-in heterozygous mice) and three testis lysates of pAcr-EGFP transgenic mice were immunoprecipitated using anti-GFP mAb magnetic beads. The GFP-IP products of six samples from two groups were subsequently analyzed by mass spectrometry (MS) to identify differentially immmunoprecipitated proteins (DIPs) in the PRSS37-EGFP group. Proteins detected in pAcr-EGFP group were used as negative controls to exclude the endogenous proteins and polypeptides that interact nonspecifically with the anti-GFP mAb magnetic beads.
Project description:Lgr5 positive and negative MaSC enriched P4 subpopulation were isolated from virgin Lgr5-EGFP-IRES-CreERT2 mice. The transcriptome profiles of the cells are compared to elucidate the molecular mechanism Lgr5+_MaSCs as a more defined MaSCs subpopulation within the P4 (MaSC and basal progenitor enriched popuation).
Project description:Data set contains samples from isolated cells of murine hearts after myocardial infarction (LAD surgery). We used an endothelial tracing system to mark endothelial cells with EGFP prior to infarct. In brief, Cdh5-ERT2+/-;mT/mG mice, which were used at an age of 10-12 weeks. Cre expression was induced by daily intraperitoneal injections of 2 mg tamoxifen for 1 week in all mice. One week after tamoxifen injection, left anterior descending coronary artery ligation surgery was performed, inducing myocardial infarction. We collected samples at homeostasis, days(d) 1, 3, 7, 14 and 28 after infarction and isolated the non-cardiomyocyte fraction for single cell sequencing. Murine scRNA-seq libraries were prepared using Chromium Single Cell 3′ v3 Reagent Kit (10X Genomics), according to manufacturer’s protocols. Raw reads were mapped against a customized version of mm10 (GRCm38.p4) containing sequences of EGFP and tdTomato.
Project description:We overexpressed Rpl22-HA in astrocytes of the mouse striatum in both wild type (WT) mice and in Crym KO mice at five months of age. Crym has been shown to be expressed predominantly in striatum and is highly expressed in astrocytes, however the function of this protein in astrocytes is unknown.
Project description:We provide an annotated cDNA clone collection which is particularly suitable for transcriptomic analysis in the mouse brain. Using it on microarrays, we compared the transcriptome of EGFP positive and negative cells in a parvalbumin-egfp transgenic background and showed that more than 30 % of clones are differentially expressed. Our clone collection will be a useful resource for the study of the transcriptome of single cell types. Keywords: Cell type comparison Comparison between fluorescent and non-fluorescent cells isolated from the visual cortex of parvalbumin-egfp transgenic mice. Using five biological replicates with a dye-swap strategy (10 hybridisations).
Project description:Alb/TGF-ß1 (TGF-ß) transgenic mice on mixed (CBAxB6) genetic background are characterized by renal fibrosis with mild or severe phenotype. Our aim was to examine the influence of genetic background on TGF-ß induced renal fibrosis in transgenic mice, and to elucidate the molecular details of strain dependent progression of fibrosis. We generated congenic B6-TGFß transgenic mice and CBAxB6-TGFß transgenic hybrid mice. Survival, proteinuria, renal histology, renal transcriptome using cDNA microarray and protein expression were analysed.