Project description:Purpose: The purpose of the study was to investigate the differential expression pattern of genes in Rag2 KO mice spleen compared to its wild type counterpart. Methods: In this study, we have excised the spleen tissues from three Rag2 KO mice and three wild type mice, and then RNA samples were prepared with the spleen tissues for the analysis of transcriptomic profiles using the Illumina MouseRef-8 v2 Expression BeadChip platforms. The MouseRef-8 v2.0 BeadChip Kit content is derived from the national center for biotechnology information reference sequence (NCBI RefSeq) database (Build 36, Release 22). The content was supplemented with probes derived from the mouse exonic evidence based oligonucleotide (MEEBO) set, as well as standard protein-coding sequences described in the RIKEN FANTOM2 database. As well, the MouseRef-8 v2.0 BeadChip targets approximately 25,600 well-annotated RefSeq transcripts, over 19,100 unique genes, and enables the interrogation of 8 samples in parallel. The MouseRef-8 v2.0 BeadChip Kit uses the DirectHyb assay and is compatible with the iScan, HiScan, and Bead array reader systems. Result: The genes expression BeadChip array analysis showed that many genes have been expressed differentially between Rag2 KO and wild type mice spleen. These genes might be involved in the different biological and physiological processes including immune regulations. Conclusion: The differential genes expression profile will provide important insight about the immune deregulation in Rag2 KO mice.
Project description:Purpose: miRNAs are important fators that are involved in the regulation of at least one third of the total human genes and are involved in the regulation of different biological processes. The purpose of the study was to investigate the differential expression pattern of miRNAs in Rag2 KO mice spleen compared to its wild type counterpart. Methods: In this study, we have excised the spleen tissues from three Rag2 KO mice and three wild type mice, and then RNA samples were prepared with the spleen tissues for the analysis of miRNAs profiles using the Affymetrix Genechip miRNA 4.0 arrays. The Affymetrix Genechip miRNA 4.0 array offers an updated content without compromising the high performance as the previous-generation arrays, it also provides a comprehensive coverage that is designed to interrogate all mature miRNA sequences in miRBase Release 20, as well as an easily correlate miRNA results having analysis files contain host gene ID, predicted and validated miRNA target genes, and clustered miRNA information. Result: The miRNAs expression microarray analysis showed that many miRNAs have been expressed differentially between Rag2 KO and wild type mice spleen. These miRNAs might be involved in the different biological and physiological processes including immune regulations. Conclusion: miRNAs might be an active player during theprocess of immune regulation.
Project description:Investigation of the SHIP1 (Inpp5d)-dependent and Rag2/Il2rg-dependent transcriptional changes in bone and osteoprogenitor cells cultures from murine strains with SHIP1 deficiency (Inpp5dm1Btlr/ m1Btlr, http://www.informatics.jax.org/allele/key/65037) and controls. Specifically, transcriptomic analysis was performed from femoral diaphysis from SHIPstyx/styx, SHIP1+/styx, wild-type (C57BL6/J), Rag2-/-/Il2rg-/-/SHIP1styx/styx, Rag2-/-/Il2rg-/-/SHIP1+/styx, and Rag2-/-/Il2rg-/-/SHIP1+/+ mice (n=3 for each genotype). Alternatively, osteoprogenitor cells isolated from bone marrow from the same genotypes as above were differentiated in vitro into primary osteoclasts in the presence or absence of RANKL. After 5 days, RNA was extracted from these cultures for transcriptomic analysis (n=3 for each genotype). Experimental Designs: Transcripts were analyzed in femoral diaphysis of SHIPstyx/styx, SHIP1+/styx, wild-type (C57BL6/J), Rag2-/-/Il2rg-/-/SHIP1styx/styx, Rag2-/-/Il2rg-/-/SHIP1+styx, and Rag2-/-/Il2rg-/-/SHIP1+/+ mice (n=3 for each genotype). Alternatively, transcripts were analyzed in primary osteoclast cultures from bone marrow cells isolated from SHIPstyx/styx, SHIP1+/styx, wild-type (C57BL6/J), Rag2-/-/Il2rg-/-/SHIP1styx/styx, Rag2-/-/Il2rg-/-/SHIP1+styx, and Rag2-/-/Il2rg-/-/SHIP1+/+ mice (n=3 for each genotype), which were supplemented or not with RANKL.
Project description:The goals of this study were to identify alterations in the gene expression profile (GEP) of spleen B cells purified from Dock10 knockout (ko) mice [C57BL/6 Dock10tm1a(EUCOMM)Hmgu/Ieg, European Mouse Mutant Archive] by comparison with the GEP of spleen B cells purified from C57BL/6 wild-type (wt) mice, and to identify alterations in the GEP of spleen B cells of the Dock10 ko mice in response to culture with IL-4 during 18 hours, by comparison with the response of the wt mice.