Project description:2 types of dendritic cells (DCs) can be generated in vitro in the presence of Flt3-L: CD4+ equivalent CD24- DCs and CD8+ equivalent CD24+ DCs. miR-142-/- mice show a severe defect in the generation of CD4+ equivalent CD24- DCs. To understand the underlying mechanism, RNA expression was analyzed by Affymetrix microarray from the 2 in vitro subtypes of DCs derived from miR-142+/+ and miR-142-/- bone marrow cells. We used microarrays to detail the global programme of gene expression in the presence or absence of miR-142 in in vitro derived DCs. Bone marrow cells from miR-142+/+ and miR-142-/- C57Bl/6 mice were isolated and incubated in the presence of Flt3-L for 8 days. in vitro derived wt and ko dendritic cells were devided into CD4+ and CD8+ equivalent DCs by FACS and sorted with a FACS-Aria. RNA was isolated and gene expression was investigated
Project description:DCs are critical for regulating immune responses. miRNAs are small, noncoding RNAs and play a significant role in regulating immune responses. miR-142 is a hematopoietic specific miRNA. To explore the role of miR-142 in regulating DCs’ immune responses, we generated mutant mice bearing a targeted deletion of the miR-142 gene. We used microarrays to detail the global program of gene expression underlying the profile changes between miR-142 KO and WT DCs and identified distinct classes of up-regulated or down-regulated genes in steady state or LPS stimulation during this process.
Project description:To gain insight into how miR-142 deficit drives a BC-like transformation, we performed RNA-seq on bone marrow (BM) Lin-Sca-1+c-Kit+ cells (LSKs) harvested from normal miR-142+/+ (wt) and miR-142−/− (miR-142 KO) mice, as well as from leukemic miR-142+/+ BCR-ABL (CP CML) and miR-142−/− BCR-ABL (BC CML) mice, two weeks after BCR-ABL induction. We then performed gene expression profiling analysis using data obtained from RNA-seq of 24 samples of LSK cells from 4 mouse strains (KO vs WT, KO CML vs CML).
Project description:2 types of dendritic cells (DCs) can be generated in vitro in the presence of Flt3-L: CD4+ equivalent CD24- DCs and CD8+ equivalent CD24+ DCs. miR-142-/- mice show a severe defect in the generation of CD4+ equivalent CD24- DCs. To understand the underlying mechanism, RNA expression was analyzed by Affymetrix microarray from the 2 in vitro subtypes of DCs derived from miR-142+/+ and miR-142-/- bone marrow cells. We used microarrays to detail the global programme of gene expression in the presence or absence of miR-142 in in vitro derived DCs.
Project description:miR-142 gene is specifically and abundantly expressed in hematopoietic cells. Mice that lack this miRNA gene develop immunodeficiency and display altered hematopoeisis. We used microarrays to detect whole transcriptome changes in miR-142 null B cells. RNA from purified WT(n=3) and miR-142 KO (n=3) CD19+ B cells was extracted and hybridized to Affymetrix GeneChips. Samples in WT and KO groups are biological replicates and were isolated in age and gender matched mice.
Project description:T cells are critical for modulating immune responses. miRNAs are small, noncoding RNAs and play a significant role in T cell responses. miR-142 is a hematopoietic specific miRNA. To explore the potential role of miR-142 in regulating T cell responses, we generated mutant mice bearing a targeted deletion of the miR-142 gene. We used microarrays to detail the global programme of gene expression underlying the profile changes between miR-142 KO and WT T cell and identified distinct classes of up-regulated genes during this process. miR-142 KO mice and WT littermates (biological triplicates) matched with age and sex were selected. T cells were purified from spleens by negative selection and processed for RNA isolation and hybridization on Affymetrix microarrays.
Project description:T cells are critical for modulating immune responses. miRNAs are small, noncoding RNAs and play a significant role in T cell responses. miR-142 is a hematopoietic specific miRNA. To explore the potential role of miR-142 in regulating T cell responses, we generated mutant mice bearing a targeted deletion of the miR-142 gene. We used microarrays to detail the global programme of gene expression underlying the profile changes between miR-142 KO and WT T cell and identified distinct classes of up-regulated genes during this process.
