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: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:This experiment was investigating how gut commensal bacteria and intestinal inflammation affect miRNA expression. We analyzed miRNA expression of spleen and intestine from specific pathogen free (SPF) B6 mice, germ-free (GF) B6 mice, and IL-10 knockout mice which have severe colitis by microarray. Thus we have total 6 samples: GF spleen; GF intestines; SPF spleen; SPF intestine; IL-10 KO spleen and IL-10 KO intestine. We directly isolated RNA from whole spleens or intestines without any treatments, and then did microarray analysis.
Project description:To understand Loxl3 gene regulation function in vivo, whole genome microarray expression profiling was used as a discovery platform to identify differential gene expressions in spleen between wild type (wt) mice and Loxl3-/- (ko) mice. Genes including cell-cell adhesion and STAT signaling pathway were identified to be differentially expressed in wt and Loxl3-/- mice.
Project description:Environmental changes may alter gene expression in depression and anxiety disorders through epigenetic regulation, including small non-coding RNAs (sncRNAs) and their major subclass, microRNAs (miRNAs). However, underlying mechanisms mediating miRNA regulation in response to changing environmental stimuli are unclear. Using the serotonin transporter (5-HTT) knockout (KO) mouse model of depression/anxiety, this study aimed to compare the effects of voluntary exercise versus chronic treatment with the stress hormone, corticosterone, on miRNA transcriptome and proteome. Using high-throughput sequencing of miRNAs and mass spectrometry (MS)-based approaches for protein expression, we described 337 differentially expressed (DE) miRNAs and 67 proteins in 5-HTT KO mice compared to wild-type (WT) control mice in standard-housing conditions. After exercise, there were 200 DE miRNAs and 3 DE proteins in WT mice, and 20 DE miRNAs and 95 DE proteins in 5-HTT KO mice, while corticosterone treatment led to 168 DE miRNAs and 1 DE protein in WT, and 21 DE miRNAs and 21 DE proteins in 5-HTT KO mice. Serotonergic dysfunction (due to the 5-HTT KO gene mutation) induced altered expression of miRNAs and proteins involved in regulation of neurodevelopment, neurogenesis and neuroinflammatory responses. Elevation of the stress hormone corticosterone activated similar (to 5-HTT KO) molecules in WT mice, while exercise caused antidepressant-like effects. We suggest that these findings indicate that functional 5-HTT might be required for the beneficial effects of exercise on miRNA expression. Our study is the first to explore how gene-environment interactions affect miRNA/proteomics composition in a mouse model of depression/anxiety and extends our understanding of gene-environmental interactions underlying these affective disorders.
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.
Project description:TAp63 is a transcription factor belonging to the p53 family with important tumor suppressive functions. We show that TAp63-/- mice exhibit an increased susceptibility to UVR-induced cutaneous squamous cell carcinoma (cuSCC). These tumors showed global disruption of miRNA and mRNA expression when compared to tumors arising in wild-type mice. A comparison to similarly sequenced human cuSCC tumors identified miR-30c-2* and miR-497 as being significantly underexpressed in cuSCC. Reintroduction of these miRNAs significantly inhibited the growth of cuSCC cell lines and xenografts. Proteomic profiling of cells transfected with either miRNA showed significant downregulation of proteins related to cell cycle progression and mitosis. A cross-platform comparison of the RNAseq and proteomics signatures identified 7 downregulated proteins, which are also frequently overexpressed in both mouse and human cuSCC. Knockdown of AURKA, KIF18B, PKMYT1, and ORC1 in cuSCC cell lines suppressed tumor cell proliferation and induced cell death. Additionally, we found that an investigational, oral, selective inhibitor of AURKA suppressed cuSCC cell growth and induced cell death, and showed anti-tumor effects in vivo. Our data establishes TAp63 as an essential regulator of miRNA expression during skin carcinogenesis and reveals a novel network of miRNAs and mRNAs, which include potential targets for therapeutic intervention.
Project description:Long non-coding RNAs (lncRNAs) play key roles in various biological processes. However, the roles of lncRNAs in macrophage polarization remain largely unexplored. Our study have found that lncRNA AK085865 play a vital role in macrophage polarization and specific bind to ILF2 protein and further influence the miRNA processing process. The miRNA expression profiles in the BMDM of C57BL/6J wild type and lncRNA AK085865 knock-out mice were obtained using microarray analysis. Based on the threshold of fold-change ≥2.0 and a p-value ≤ 0.05, we screened a total of 24 miRNAs, of which 11 were upregulated and 13 were downregulated in KO compared to that in WT BMDMs.This study provides new insights into the role of lncRNA AK085865 regulating miRNAs expression in the mice BMDMs.
Project description:To examine the effect of DEAD-box RNA helicase subunits on the primary miRNAs processing by Drosha complex, we made knockout mice of p72, DEAD-box RNA helicase, a component of Drosha complex. And we compare the miRNA expression profiles derived from whole mice embryo between wild-type and p72 KO mice.