Conjoint analysis of RNA-seq and Solexa sequencing data of bovine mammary epithelial cells isolated from Chinese Holstein cows with high and low milk fat percentages
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ABSTRACT: For RNA-seq, the sequencing library of each RNA sample was prepared by using Ion Total RNA-Seq Kit v2. For Solexa suquencing, we download the data obtained by a previous study in our laboratory. Genes and miRNAs were considered differentially expressed according to log2FC >1 or log2FC <-1and FDR <0.05. All target genes regulated by DERs were predicted by the combination of miRbase (http://www.mirbase.org) and TargetScan (http://www.targetscan.org). All the predicted target genes were intersected with the DEGs obtained by RNA-seq, and then the DEGs and DERs were obtained by a combined analysis.In total, 190 DEGs were regulated by 33 DERs that were significantly enriched in 488 GO terms and 12 KEGG pathways (p <0.05).This is of great significance for research on miRNAs and functional genes for the milk fat percentage- and marker-assisted selection of dairy cattle.
Project description:1. The expression of plasma exosomal ncRNAs and mRNAs was compared a group of 12 samples from healthy individuals (normal group) with a group of 30 samples from cervical cancer patients before primary concurrent chemoradiotherpay (cancer group) using differentially expressed gene (DEG) analysis. The RNAs with both |log2FC| >2 and P values < 0.05 2. We prepared for log2 fold change values of plasma exosomal RNAs for 30 patients from 30 samples in second week during CCRT compared with those before treatment to screen secondary biological function of primarily selected DEGs. We analyzed the association between non-coding RNA (ncRNA)-mRNA network and cancer using ingenuity pathway analysis after secondary selection according to the number and correlation of mRNAs (or ncRNAs) relevant to log2FC in the expression of primarily selected ncRNAs (or mRNAs) before and after CCRT. The raw NGS data of 58 samples from 29 patients were uploaded in E-MTAB-10215. Here we will upload raw RNA sequencing data of 12 samples from 12 healthy individuals and 2 samples from one cervical cancer patient.
Project description:Background: Bone nonunion is a serious complication of fracture. This study explored the differentially expressed lncRNAs (DELs) and mRNAs (DEGs) and identified potential lncRNA-mRNA interactions in bone nonunion. Methods: We extracted total RNA from three bone nonunion and three bone union patient tissue samples. RNA sequencing was performed to detect DELs and DEGs between bone nonunion and union tissue samples. The lncRNAs and genes with absolute log2-fold change (log2FC) > 1 and adjusted p value < 0.05 were further chosen for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. lncRNA and targeted mRNA interaction networks were constructed. Results: We observed 179 DELs and 415 DEGs between the bone nonunion and union tissue samples. GO analysis indicated that DELs and DEGs were mainly enriched in chondroitin sulfate proteoglycan biosynthetic process. DELs and DEGs were enriched in 'ECM-receptor interaction' and 'Staphylococcus aureus infection' KEGG pathways. Several potential lncRNA-mRNA interactions were also predicted. Conclusions: This study identified bone nonunion-associated lncRNAs and mRNAs using deep sequencing that may be useful as potential biomarkers for bone nonunion.
Project description:Radiation-induced pulmonary fibrosis (RIPF) is the most common bottleneck of radiation therapy (RT) that limits its tumor-killing effect. Up to date, there is no effective therapeutic agent for RIPF treatment due to absence of realistic preclinical models. Lungs of mice exposed to focal, highly localized dose of 75Gy-100Gy represents short time-taking (up to 6 weeks) modality for mimicking recent advanced RT, stereotactic body RT-associated lung fibrosis. However, extensive fibrosis and possibility of animal lethality resulted from exposure to high dose raises a question regarding the appropriate dose threshold for limiting unnecessary responses and obtain clinically relevant results. We observed in preliminary trials that lung tissues of 65Gy-exposed mice are still able to develop inflammation and fibrosis at the same time cap. But the molecular changes associated with dose reduction is still unknown. To examine this, bulk RNA from 65Gy and 75Gy irradiated as well as normal lung tissues of mice were obtained, and analysed using computational approach to uncover the transcriptomal similarities and differences between the differentially irradiated samples. Differentially expressed genes (DEGs) were analysed and 2-week and 6-week intersected DEGs as well as 65 and 75Gy exclusive DEGs at 2-week and 6-week time points were detected using venn diagramme. Expression direction of intersected DEGs between 65Gy and 75Gy was checked and found to be consistent at both inflammation and fibrosis stages proven visually by MDS and heatmap graphs. All-times intersected DEGs are 488 and involved in inflammation, cytokine production, and cell proliferation; 2-week intersected DEGs are 221 and involved in cell cycle and DNA damage; 6-week intersected DEGs are 403 and involved in immunoglobin production, extracellular matrix (ECM) deposition, and ECM-receptor interactions. Interestingly, only 75Gy exclusive DEGs of 2 and 6 weeks presented inflammation and fibrosis-related biological process, respectively. These include anti-inflammatory process, cytokine production, EMT, vasculogenesis, and angiogenesis. These results suggested that 65Gy maybe a better dose for delivering less exaggerated, clinically relevant inflammation and fibrotic events, and thus, it can be utilised in establishing appropriate preclinical models for studying RIPF pathology and therapy.
Project description:This study aim to screening potential regulators of fecundity, the bovine testes of immature and mature in Chinese Red Steppes were performed by a genome-wide analysis of mRNAs and miRNAs. Compared with born testicular tissue, 6051 upregulated genes and 7104 downregulated genes in adult bovine testicular tissue were identified as differentially expressed genes (DEGs) (log2FC>1 or <-1 , FDR <0.05). The DEGs were significantly enriched in 808 GO terms (p <0.05), which are involved in sperm biological activity, including male gonad development, male genitalia development, spermatogenesis, sperm motility, spermatid development, sperm chromatin condensation, etc. Moreover, DEGs were also significantly enriched in 105 KEGG pathways (p <0.05), such as focal adhesion, cGMP-PKG signaling pathway and calcium signaling pathway, etc. To explore the miRNA-regulated gene expression network integrated analysis was used between DEGs and differentially expressed miRNAs (DERs). Correlation prediction and analysis found that 896 differentially expressed target genes were negatively correlated with the expression levels of 31 differentially expressed bovine miRNAs. Our results identified novel candidate DEGs and DERs correlated with male reproduction, which will be valuable for future genetic and epigenetic studies of sperm development and maturity, as well as provided valuable insights into the molecular mechanisms of the male fertility and spermatogenesis in cattle.
Project description:We used an RNA-Seq transcriptomic approach to investigate Longissimus dorsi muscle gene expression profiles of calves from cows receiving weekly Se-yeast boluses at supranutritional concentrations during different trimesters of gestation. Pregnant beef cows received, except the control group (CTR), weekly supranutritional selenium-yeast boluses (105 mg Se/wk) during the first (TR1), second (TR2), or third (TR3) trimester of gestation. Within 12 to 48 h of birth, muscle samples were collected from the Longissimus dorsi using a Bergstrom biopsy needle (inner diameter: 5 mm). Muscle biopsies were placed into sterile vials, stored on ice and frozen at – 80 ºC until total RNA isolation was performed. After sequencing and read quality control, we identified 3,048 unique differentially expressed genes (DEGs) across all group comparisons (FDR < 0.05 and |log2FC| > 1.5). Furthermore, we predicted 237 unique transcription factors that putatively regulate the DEGs. Our findings suggest a beneficial effect of supranutritional maternal organic Se supplementation during late gestation on Se-status and muscle development and function of newborn calves.
Project description:Purpose and methods:Transcriptome profiling of Phytophthora sojae P6497 mycelium (3-days old) and Phytophthora infestans T30-4 mycelium (6-days old) were generated to find out the relationship between 6mA methylation and gene expression. RNA-seq data was mapped using Tophat2, and gene expression data was generated by Cufflinks. Transcriptome profiling of P. sojae psdamt3 mutant T9 (lost 374bp by CRISPR/Cas9) was generated to check the differential expressed genes (DEGs) between the mutant and wild-type P. sojae P6497. Read counts was calculated using featureCounts, and DEGs was calculated using DEseq2 with |log2FC|≥1, y-axis is FDR<0.05. Conclusions: 6mA is associated with lowly expressed genes. Examination of differentially expressed genes (DEGs) in psdamt3 uncovers a total of 3156 genes, with 1544 genes up-regulated and 1622 genes down-regulated.
Project description:The goal of this study was to compare differentially expressed transcripts in onion bulbs induced in response to managed stress under field conditions using transcriptome profiling (RNA-seq). The clean reads were aligned to the reference genome (a doubled haploid onion line DHCU066619 (https://www.oniongenome.wur.nl/) and the transcriptions assembly. The read count for every transcript was determined from the findings of the mapping process. Genes identified as differentially expressed genes (DEGs) had an adjusted P-value <0.05 and an absolute log2FC ≥0, as determined by DESeq2. Transcriptomic analysis of onion bulb tissue (variety Matahari) in response to nitrogen stress under optimal water identified 3092 and 2204 Differentially Expressed Genes (DEGs) unique genes under optimal nitrogen vs low nitrogen. while under sub-optimal nitrogen, 2770 and 2198 unique genes were identified under optimal nitrogen vs low nitrogen respectively.
Project description:The anthocyanin glycosides in potato tubers not only provide the plant with bright color and high nutritional value, but also have pharmacological effects such as antioxidant, hypotensive, lipid-lowering and immunity-boosting. However, the regulatory mechanism of anthocyanin in potato tubers is still unclear. In this study, the content of anthocyanin was measured in four potato varieties (B, R, Y and W) and genes regulating anthocyanin synthesis in different potato varieties were identified by comparing transcriptomic approaches. The results showed that 82.6G of filtered data were obtained in 12 sequencing libraries, of which 82.23-87.66% of clean fragments were covered to the reference genome. In total, 42,793 genes were found in the twelve libraries of which 3765 were new genes. Finally, 3638 (B), 4295 (R) and 3594 (Y) DEGs were obtained in the three groups by the strict threshold screening (FDR<0.05 and |log2FC|>1), respectively. GO analysis showed that DEGs were mainly enriched in entries such as cell (GO:0005623), metabolic process (GO:0008152), catalytic activity (GO:0003824), etc. KEGG analysis revealed that metabolic pathways (ko01100), phenylalanine metabolism (ko00360), photosynthesis (ko00195) were significantly enriched. In addition, some genes related to anthocyanin synthesis (e.g. CHS, bHLH and MYB) were highly induced or repressed in different varieties. In conclusion, the results of this study provide insight into the genes regulating anthocyanin formation in potato and provide a scientific reference for breeding better quality potato varieties.
Project description:In this study, we aimed to explore RNAs profiling regulated by LNPPS in bladder cancer. RNA-seq analysis of three pairs of LNPPS-overexpressing 5637 cells and control cells was completed in this project. A total of 17.6Gb raw data was obtained. In concluion, our data showed that the overexpression of LNPPS caused the significantly upregulation of 82 mRNAs, and the downregulation of 102 mRNAs (|Log2FC|>1.5, p<0.05). Further studies will focus on the potential regulatory mechanisms of these significantly differently expressed mRNAs in the LNPPS-mediated development of bladder cancer.
Project description:To further investigate the therapeutic potential of NBXH in treating tuberculosis, we employ whole genome microarray expression profiling as a discovery platform. The study aimed to explore the impact of NBXH treatment on the gene expression profile of tuberculosis-infected animal models. The animal models were treated with high, medium, and low doses of NBXH. Following three months of treatment, PBMCs from each treatment group were used for whole genome microarray expression profiling analysis, alongside a control group. With a screening criterion of P <0.05 and an absolute log2FC value >1, 3454 up-regulated and 3597 down-regulated DEGs were respectively identified in the gene expression profiles of the mouse TB model treated with NBXH.