Project description:To determine the circRNA expression profile in preeclampsia and natural pregnancy placenta tissues, we uesed circRNA microArray analysis form Arraystar to examine the expression of circRNAs in preeclampsia and natural pregnancy placenta tissues.
Project description:To investigate the expression profiles of circRNAs in the placental tissues from pregnant women with fetal growth restriction (FGR), Arraystar Human circRNA Array V2 analysis was performed. Six samples (three samples each from FGR group and healthy controls group) were used in the microarray screening. 13532 circRNAs were identified in the placental tissues from FGR and normal groups, and only 244 circRNAs were found to be more differentially expressed in the placental tissues of the FGR group than in normal placenta (fold-change > 1.5, p < 0.05). Among them, 100 and 144 circRNAs were up- and down-regulated, respectively. To verify circRNA expression in the FGR placenta, we chose hsa_circ_0007738, hsa_circ_0071271, and hsa_circ_0000848, which have the greatest expression differences between FGR and normal groups, for further analysis. Hsa_circ_0000848 expression level was significantly lower in FGR placental tissues than in the normal placenta tissues, whereas hsa_circ_0007738 and hsa_circ_0071271 expression levels did not change significantly. Further, we found that hsa_circ_0000848 promoted trophoblast cell migration and invasion, and inhibited cell apoptosis via sponging miR-6768-5p.
Project description:To better study the bovine circRNA, we assessed the outcomes of circRNA identification using six enrichment approaches. RNA-sequencing analysis revealed that different approaches led to varied the ratio of uniquely mapped reads, false positive rate of identifying circRNAs, and the number of circRNAs per million clean reads. Besides, 507 of 4,051 identified bovine high confident circRNAs had shared splicing sites with human circRNAs.
Project description:Gene expression patterns of bovine placenta from initial to late stage of pregnancy were investigated by using the our custom designed utero-placental cDNA microarray. Keywords: Time course
Project description:MicroRNA (miRNA) sponges containing miRNA complementary binding sites constitute a potentially useful strategy for miRNA-inhibition therapeutics in cancer patients. Recently, naturally occurring circular RNAs (circRNAs) have been revealed to function as efficient microRNA sponges. We hypothesized that synthetic circRNA sponges targeting oncomiRs could be constructed and used to achieve potentially therapeutic microRNA loss of function. In this study, linear RNA molecules containing five miR-21 binding sites were transcribed in vitro. After dephosphorylation by calf intestinal phosphatase and phosphorylation by T4 polynucleotide kinase, circRNA sponges were circularized using 5’-3’ end ligation by T4 RNA ligase 1. Synthetic circular sponge stability was assayed in the presence of RNase R or fetal bovine serum. Luciferase reporter and cell proliferation assays were performed to assess competitive inhibition of miR-21 activity by circRNA sponges in NCI-N87 gastric cancer cells. Tandem Mass Tag (TMT) labeling proteomics analysis and Western blotting were performed to delineate effects of circRNA sponges on miR-21 downstream targeted proteins. Our experiments revealed that artificial circRNA sponges can be synthesized using enzymatic ligation. These synthetic circRNA sponges are more resistant than their linear RNA counterparts to nuclease degradation in vitro. They effectively suppress the activity of miR-21 on its downstream protein targets, including the important cancer protein DAXX. Finally, they also inhibit gastric cancer cell proliferation. Our results suggest that synthetic circRNA sponges represent a rapid, effective, convenient strategy to achieve loss of miRNA function in vitro, with potential future therapeutic application in vivo.
Project description:RNA sequencing was performed to obtain a landscape of mRNA, circRNA, lncRNA, and miRNA of bovine intramuscular adipocytes in 4 differentiation periods.
Project description:Purpose: to detect expression profile of RNA (lncRNA and circRNA) and elucidate differentially expressed RNAs (DElncRNAs and DEcircRNAs) with potential roles during lipopolysaccharide (LPS)-induced inflammation models of bovine mammary epithelial cells MAC-T in vitro. Methods: bovine mammary epithelial cells MAC-T were exposed to LPS for 0, 6 and 12 hours to assess the expression profiles of RNA (lncRNA and circRNA) using RNA-seq. Results: totally 112 DElncRNAs and 71 DEcircRNAs were screened out at different time points. Functional enrichment analysis on target genes of lncRNAs and host genes of circRNAs indicated that these genes were involve in regulating inflammation-related signaling pathways, including Notch, NF-κB, MAPK, PI3K-Akt, mTOR, MAPK and NOD-like receptor signaling pathway. Conclusion: these differentially expressed RNAs (DElncRNAs and DEcircRNAs) may be involved in the regulation of a variety of immune-related processes including inflammatory responses bovine mammary epithelial cells exposed to LPS via some vital signaling pathways. This study lays a foundation for further research on molecular regulation of bovine mastitis, and also provides a reference for breeding strategies based on molecular markers for mastitis resistance in dairy cows.
Project description:Ruminants have a semi-invasive placenta, which possess highly vascularized placentomes formed by maternal endometrial caruncles and fetal placental cotyledons and required for fetal development to term. The synepitheliochorial placenta of cattle contains at least two trophoblast cell populations, including uninucleate (UNC) and binucleate (BNC) cells that are most abundant in the cotyledonary chorion of the placentomes. The interplacentomal placenta is more epitheliochorial in nature with the chorion developing specialized areolae over the openings of uterine glands. Of note, the cell types in the placenta and cellular and molecular mechanisms governing trophoblast differentiation and function are little understood in ruminants. To fill this knowledge gap, the cotyledonary and intercotyledonary areas of the mature day 195 bovine placenta were analyzed by single nuclei analysis. Single-nuclei RNA-seq analysis found substantial differences in cell type composition and transcriptional profiles between the two distinct regions of the placenta. Based on clustering and cell marker gene expression, five different trophoblast cell types were identified in the chorion, including proliferating and differentiating UNC and two different types of BNC in the cotyledon. Cell trajectory analyses provided a framework for understanding the differentiation of trophoblast UNC into BNC. The upstream transcription factor binding analysis of differentially expressed genes identified a candidate set of regulator factors and genes regulating trophoblast differentiation. This foundational information is useful to discover essential biological pathways underpinning the development and function of the bovine placenta.