Screening of influenza A virus infection related microRNA
Ontology highlight
ABSTRACT: In the study presented here, A549 cells infected with influenza virus A /JingFang/86-1(H1N1) for 4 and 24 hours and intervention by Flutide, was used to acquire expression profiles of a total of 471 standard mature miRNAs based on Sanger miRBase Release 9.2. 4 samples: uninfected, 4 h and 24 h after infection, and 24 h infection then treated with Flutide
Project description:We detect the small RNAs subcellular distribution in breast cancer cell lines MCF-7 and MDA-MB-231, and normal cell line MCF-10A. Each cell line, we detected the nuclear and cytoplasmic small RNAs expression intensity; and then we could get the nuclear-cytoplasmic-ratio.
Project description:Cardiac profiling of miR expression levels in a transgenic mouse model of heart failure (MHC-CnA) to identify miRs that are co-regulated with the development of calcineurin-induced heart failure. Two-condition experiment: WT vs CnTg. Biological replicates: one array was performed on 3 pooled samples of each genotype (2 arrays in total).
Project description:In this study, we examined if the composition of plasma miRNAs is altered in patients with traumatic brain injury (TBI), and if these changes can be used as diagnostic markers. A microarray containing 875 human miRNAs was used to compare the miRNA profile of plasma collected from severe TBI patients (GCS M-bM-^IM-$ 8) to that of age-, gender-, and race-matched healthy volunteers. This screen identified 108 miRNAs in the plasma of healthy volunteers. Of these, 52 were found to be altered in plasma samples from persons with severe TBI, and an additional 8 miRNAs were detected only in the plasma of TBI patients. Plasma samples from 10 patients from either severe TBI (experimental group) or healthy volunteers (reference group; age-, gender-, and race-matched ) were pooled, the total RNA extracted in parallel, eluted in 100ul, and dried to 30 ul. Equal volumes of extracted plasma RNAs were assayed for global miRNA content using a service provider (LC Sciences, Houston, TX). There were no replicates performed for this screen. Healthy volunteer group served as the reference.
Project description:In this study, miRNA expression in splenic lymphocytes from three genetically disparate lupus-prone mouse models (MRL-lpr, B6-lpr and NZB/WF1) were profiled. 49 miRNAs were found to be differentially expressed in MRL-lpr mice compared to MRL mice; and 24 miRNAs were differentially expressed in B6-lpr mice compared to B6 mice. Among these dysregulated miRNAs, we noted that 15 miRNAs were common to both lpr strains. Interestingly, microarray analysis of NZB/W and NZW at 3 months of age, an age when overt lupus disease is not evident in NZB/W mice, revealed that only one miRNA, miR-148a was significantly upregulated in NZB/W mice. The aim of this porject is to determine the common miRNA expression changes in splenocytes from different strains of murine lupus models. The splenocytes were prepared from genetically lupus-prone female mice including MRL/MpJ-Faslpr/J (MRL-lpr), NZBWF1/J (NZB/W), B6.MRL-Faslpr/J (B6-lpr) and their control mice MRL/MpJ (MRL), NZW/LacJ (NZW) and C57BL/6J (B6) mice (The Jackson laboratory, ME). Total RNAs, containing miRNAs were isolated from whole splenocytes using mirVana miRNA isolation kits (Ambion) following manufactory’s instructions and sent to LC Sciences (http://www.lcsciences.com/) for the microarray assay. The mouse miRNA array chips (Chip ID miRMouse 12.0 version), which included 617 unique, mature, mouse miRNA, based on the Sanger miRBase Release 12.0, were used in the assay.
Project description:miRNA profiling of kidney tissue from C57BL/6 mice that received a 30 minute ischemic injury compared with control kidney tissue from mice that received sham operation only. Two condition experiment - sham and ishemic injury (IRI). Eight time points were measured using pooled samples from three mice.
Project description:MicroRNAs (miRNAs), approximately 22-nucleotide non-coding RNA molecules, regulate a variety of pivotal physiological or pathological processes, including embryonic development and tumorigenesis. To obtain comprehensive expression profiles of miRNAs in human embryos, we characterized miRNA expression in weeks 4-6 of human embryonic development using miRNA microarrays and identified 50 human-embryo-specific miRNAs (HES-miRNAs). Furthermore, we selected three non-conserved or primate-specific miRNAs, hsa-miR-638, -720, and -1280, and examined their expression levels in various normal and tumor tissues. The results show that expression of most miRNAs is extremely low during early human embryonic development. In addition, the expression of some non-conserved or primate-specific miRNAs is significantly different between tumor and the corresponding normal tissue samples, suggesting that the miRNAs are closely related to the pathological processes of various tumors. This study presents the first comprehensive overview of miRNA expression during human embryonic development and offers immediate evidence of the relationship between human early embryonic development and tumorigenesis. Total 10 samples were used microRNAs microarrays hybridization, then the original data from 8 of 10 samples was used final data analysis. The samples ZN18, ZN46/47 and ZN75 were designated as Week 4 of human embryonic development; the samples ZN38, ZN43 and ZN63-1 were designated as Week 5 of human embryonic development; the samples ZN61 and ZN70 were designated as Week 6 of human embryonic development.
Project description:miRNA profiling of resting and activated T cells Two condition experiment, resting versus activated T cells, measured pooled samples from three independent stimulations
Project description:The extraocular muscles (EOMs) are a unique group of muscles that are anatomically and physiologically distinct from other skeletal muscles. Previously, we and others have shown that EOMs have a unique transcriptome and proteome. Here, we investigated the expression pattern of microRNAs (miRNAs) in EOM, as they may play a role in generating the unique EOM allotype. We screened LC Sciences miRNA microarrays covering the sequences of miRBase 10.0 to define the microRNAome of normal mouse EOM and tibialis anterior (TA) limb muscle. 74 miRNAs were found to be differentially regulated (p-value < 0.05) and 31 miRNAs (14 up-regulated and 17 down-regulated) were found to be differentially regulated at a signal strength > 500 including the muscle-specific miR-206, miR-1, miR-133a, miR-133b and miR-499. qPCR analysis was used to validate the differential expression. Bioinformatic tools were used to identify potential miRNA-mRNA-protein interactions and integrate data with previous transcriptome and proteomic profiling data. Luciferase assays using co-transfection of precursor miRNAs (pre-miRNAs) along with reporter constructs containing the 3â-untranslated region (3âUTR) of their predicted target genes were used to validate targeting by identified miRNAs. The definition of the EOM microRNAome complements existing transcriptome and proteome data about the molecular make-up of EOM and provides further insight into regulation of muscle genes. These data will also help to further explain the unique EOM muscle allotype and its differential sensitivity to diseases such as Duchenne's muscular dystrophy (DMD) and may assist in development of therapeutic strategies. Total RNA from four EOM and four TA tissue samples dissected from four adult male C57/Bl10 mice were used (TA served as control) to screen four LC Sciences microRNA Microarray chips. The chips contained microRNA sequences based on miRBase content 10.0 totalling 568 different miRNAs. Samples were labelled with Cy3 and Cy5 using dye-swap. Relative differences of miRNA expression was expressed as fold-changes EOM/TA, which were calculated after normalization across all four arrays.
Project description:Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) is a tumor suppressor that negatively regulates cell survival and proliferation by antagonizing phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PKB/Akt) signaling. Loss of heterozygosity (LOH) of PTEN, reduced expression of PTEN and overexpression of phosphorylated Akt are frequently found in human gastric cancer, and their changes correlate with tumor progression and prognosis. Previous studies have shown that the deregulated miRNAs in human gastric cancer play important roles in gastric cancer cell proliferation, apoptosis and inflammation. However, miRNAs downstream PTEN/Akt signaling is poorly investigated. To clarify whether PTEN is involved in gastric tumorigenesis, we have generated a gastric epithelium specific PTEN knockout mouse which exhibited gastric tumor formation with enhanced cell proliferation.So the objectives of the microarray experiment were to, a) screen miRNAs which might be regulated by PTEN/Akt signaling by comparing the miRNA expression profiles between PTEN deficient and control gastric epithelia. 2) explore the microRNA mechanism involved in gastric cell proliferation and gastric tumorigenesis. miRNA profiling of mouse gastric epithelium,comparing Pten mutant mouse with controls. 4 samples. Experiments in 2 different time point, 20 days and 60 days after birth, 2 Biological replicates. Mutant tissue vs. controls from mixture of 3-4 mouse.
Project description:Expression phospho-mimic TRBP enhanced in vitro miRNA production and cellular expression of ectopic miRNA. We used miRNA microarray experiments to determine the effect of expression of phopsho-mimic TRBP on global endogenous miRNA expression. We established isogenic cell lines stably expressing phospho-mutant and phospho-mimic TRBP using Flp-In 293 cells and a Flp Recombinase Target system. After monoclonal cell colonies were selected, cells were cultured for two weeks. Total RNA was harvested when cells were ~75% confluent. Initial analysis was performed using one sample from each experimental group. Subsequent analysis included an additional two samples for each group. miRNA with values less than 500 were not included in analysis.