Project description:To analyze gene expression in in regulatory T cell precursors that develop in the absence of a functional Foxp3 protein as compared to that of normal regulatory T cells Keywords: Cell type comparison
Project description:To analyze gene expression in in regulatory T cell precursors that develop in the absence of a functional Foxp3 protein as compared to that of normal regulatory T cells Experiment Overall Design: Murine wild-type and mutant (delta) Foxp3 alleles were tagged with EGFP reporters. CD4 positive T cells expressing the respective tagged Foxp3 allele (Foxp3/EGFP and delta Foxp3/EGFP) were isolated from hemizygous male mice by cell sorting using FACS. CD4 positive cells that did not express the EGFP tag were a isolated. Total RNA was prepared and used in the array studies.
Project description:The gene expression profile of peripheral Foxp3+ natural regulatory T cells isolated from Foxp3/EGFP bicistronic mice was compared to that of in vitro-induced regulatory T cells and to CD4+ conventional (Foxp3-) T cells. The role of the regulatory T cell transcription factor Foxp3 in shaping the transcriptosomes of natural and induced regulatory T cells was analyzed using mice expressing a mutant FOXP3-EGFP fusion protein (Foxp3deltaEGFP). We used gene expression microarrays to examine the transcriptional programs of natural and induced regulatory T cells and the function of Foxp3 in organizing the transcriptosomes of the respective cell type Experiment Overall Design: Conventional T cells and natural and induced regulatory T cells were derived from Foxp3/EGFP bicistronic mice and analyzed for their gene expression profile. Conventional T cells, regulatory T cell precursors (CD4+Foxp3deltaEGFP+) and induced regulatory T cell precursors (CD4+Foxp3deltaEGFP+) cells were deriv ed from Foxp3deltaEGFP mice
Project description:Analysis of Foxp3 ablated peripheral regulatory T cells. Regulatory T cells require the expression of the transcription factor Foxp3 for thymic development. It is not known whether continuous expression of Foxp3 is required for the maintained function of mature regulatory T cells in the periphery. Results indicate changes to the regulatory T cell developmental program in the absence of Foxp3. Experiment Overall Design: Compare Cre recombinase treated peripheral regulatory T cells possessing a Cre sensitive Foxp3 locus to Cre treated regulatory T cells with a wild type Foxp3 locus. Cre exposure is observed via the Cre sensitive expression of the yellow flourescent protein molecule.
