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Screening and validation of lncRNAs and circRNAs as miRNA sponges [cell lines, miRNA-2_0]

ABSTRACT: Intensive research in past two decades has uncovered the presence and importance of noncoding RNAs (ncRNAs), which includes microRNAs (miRs) and long ncRNAs (lncRNAs). These two classes of ncRNAs interact to a certain extent, as some lncRNAs bind to miRs to sequester them. Such lncRNAs are collectively called 'competing endogenous RNAs' or 'miRNA sponges'. In this study, we screened for lncRNAs that may act as miRNA sponges using the publicly available data sets and databases. To uncover the roles of miRNA sponges, loss-of-function experiments were conducted, which revealed the biological roles as miRNA sponges. LINC00324 is important for the cell survival by binding to miR-615-5p leading to the de-repression of its target BTG2 LOC400043 controls several biological functions via sequestering miR-28-3p and miR-96-5p, thereby changing the expressions of transcriptional regulators. Finally, we also screened for circular RNAs (circRNAs) that may function as miRNA sponges. The results were negative at least for the selected circRNAs in this study. In conclusion, miRNA sponges can be identified by applying a series of bioinformatics techniques and validated with biological experiments.

ORGANISM(S): synthetic construct Homo sapiens

PROVIDER: GSE73690 | GEO | 2016/07/06

SECONDARY ACCESSION(S): PRJNA297675

REPOSITORIES: GEO

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Screening and validation of lncRNAs and circRNAs as miRNA sponges [cell lines, miRNA-2_0]

Project description:Intensive research in past two decades has uncovered the presence and importance of noncoding RNAs (ncRNAs), which includes microRNAs (miRs) and long ncRNAs (lncRNAs). These two classes of ncRNAs interact to a certain extent, as some lncRNAs bind to miRs to sequester them. Such lncRNAs are collectively called 'competing endogenous RNAs' or 'miRNA sponges'. In this study, we screened for lncRNAs that may act as miRNA sponges using the publicly available data sets and databases. To uncover the roles of miRNA sponges, loss-of-function experiments were conducted, which revealed the biological roles as miRNA sponges. LINC00324 is important for the cell survival by binding to miR-615-5p leading to the de-repression of its target BTG2 LOC400043 controls several biological functions via sequestering miR-28-3p and miR-96-5p, thereby changing the expressions of transcriptional regulators. Finally, we also screened for circular RNAs (circRNAs) that may function as miRNA sponges. The results were negative at least for the selected circRNAs in this study. In conclusion, miRNA sponges can be identified by applying a series of bioinformatics techniques and validated with biological experiments. The expressions of miRs were examined in HEK-293, HUVEC and Hs68 cells.

2016-07-06 | E-GEOD-73690 | biostudies-arrayexpress

Screening and validation of lncRNAs and circRNAs as miRNA sponges [siRNA, miRNA-2_0]

2016-07-06 | E-GEOD-73689 | biostudies-arrayexpress

Screening and validation of lncRNAs and circRNAs as miRNA sponges [siRNA, HuGene-1_0-st]

Project description:Intensive research in past two decades has uncovered the presence and importance of noncoding RNAs (ncRNAs), which includes microRNAs (miRs) and long ncRNAs (lncRNAs). These two classes of ncRNAs interact to a certain extent, as some lncRNAs bind to miRs to sequester them. Such lncRNAs are collectively called 'competing endogenous RNAs' or 'miRNA sponges'. In this study, we screened for lncRNAs that may act as miRNA sponges using the publicly available data sets and databases. To uncover the roles of miRNA sponges, loss-of-function experiments were conducted, which revealed the biological roles as miRNA sponges. LINC00324 is important for the cell survival by binding to miR-615-5p leading to the de-repression of its target BTG2 LOC400043 controls several biological functions via sequestering miR-28-3p and miR-96-5p, thereby changing the expressions of transcriptional regulators. Finally, we also screened for circular RNAs (circRNAs) that may function as miRNA sponges. The results were negative at least for the selected circRNAs in this study. In conclusion, miRNA sponges can be identified by applying a series of bioinformatics techniques and validated with biological experiments. Using HEK-293 cells, we performed microarray experiments (n=2) for siRNA-mediated silencing of LINC00324 and LOC40043.

2016-07-06 | E-GEOD-73683 | biostudies-arrayexpress

Screening and validation of lncRNAs and circRNAs as miRNA sponges [siRNA, miRNA-2_0]

2016-07-06 | GSE73689 | GEO

Screening and validation of lncRNAs and circRNAs as miRNA sponges [siRNA, HuGene-1_0-st]

2016-07-06 | GSE73683 | GEO

Synthetic circular RNA functions as a miR-21 sponge to suppress gastric carcinoma cell proliferation

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.

2018-10-22 | PXD008584 | Pride

Characterization and Analysis of Whole Transcriptome of Ningxiang Pig Spleens Reveal Potential Mechanism for Transcriptional Regulation in Immunity [RNA-Seq]

Project description:we comprehensively analyzed the whole transcriptome of the spleen from three important developmental nodes(30d, 90d and 210d) of Ningxiang pig (a Chinese indigenous breed). A total of 812, 364 and 865 differentially expressed (DE) mRNAs, 98, 37 and 97 DE miRNAs, 278, 220 and 271 DE lncRNAs, and 113, 96 and 96 DE circRNAs were identified between 90d and 30d, 210d and 30d, and 210d and 90d, respectively. And their molecular characterization, spatio temporal expression pattern, potential function and interaction with miRNAs were analyzed. Additionally, co-expression network analysis identified the potential non coding RNAs(ncRNAs) related to immunomodulation, e.g., ssc-miRNA-150, ssc-miRNA-497, MSTRG.24160, MSTRG.18646. Furthermore, our results also revealed that miRNAs and circRNAs may have evolved to regulate a large set of biological processes of Ningxiang pigs spleens, and circRNAs may play the role of miRNA sponges. Our study provides new insights into the expression changes of RNAs during the spleen development process of Chinese indigenous breed Ningxiang pigs, which might contribute to the phenotypic formation of immune function in Chinese native pig breeds. These results greatly improved our understanding of the molecular mechanisms which regulate the immune function of the spleen of Ningxiang pigs.

2022-06-06 | GSE152643 | GEO

Characterization and Analysis of Whole Transcriptome of Ningxiang Pig Spleens Reveal Potential Mechanism for Transcriptional Regulation in Immunity [miRNA-Seq]

2022-06-06 | GSE152087 | GEO

circRNA, lncRNA and mRNA profiles of umbilical cord blood exosomes from preterm newborns showing bronchopulmonary

Project description:Purpose: The purpose of this study was to detect the expression profiles of circRNAs, lncRNAs and mRNAs in umbilical cord blood (UCB) exosomes from BPD infants. Methods:Microarray analysis was performed to compare the RNA profiles of UCB-derived exosomes from preterm newborn with and without BPD. Then, circRNA/lncRNA-miRNA-mRNA co-expression networks were built to determine their associations with BPD.Results:A total of 317 circRNAs, 104 lncRNAs, 135 mRNAs showed significant differential expression in UCB-derived exosomes from BPD preterm infants compared with the NBPD group. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were conducted for examining differentially expressed exosomal circRNAs, lncRNAs and mRNAs. Our results showed the GO terms and KEGG pathways mostly involving differentially expressed exosomal RNAs were closely associated with endothelial or epithelial cell development. In addition, two networks were identified, including circRNA-circ_0086018/miR-762/MEN1 and lncRNA-BASP1-AS1/miR-20a-5p/DNAJC22 networks, which may take part in BPD.

2022-08-31 | GSE190215 | GEO

Expanded identification and characterization of mammalian circular RNAs

Project description:The recent reports of two circular RNAs (circRNAs) with strong potential to act as microRNA (miRNA) sponges suggest that circRNAs might play important roles in regulating gene expression. However, the global properties of circRNAs are not well understood. We developed a computational pipeline to identify circRNAs and quantify their relative abundance from RNA-seq data. Applying this pipeline to a large set of non-poly(A)-selected RNA-seq data from the ENCODE project, we annotated 7,112 human circRNAs that were estimated to comprise at least 10% of the transcripts accumulating from their loci. Most circRNAs are expressed in only a few cell types and at low abundance, but they are no more cell-type–specific than are mRNAs with similar overall expression levels. Although most circRNAs overlap protein-coding sequences, ribosome profiling provides no evidence for their translation. We also annotated 635 mouse circRNAs, and although 20% of them are orthologous to human circRNAs, the sequence conservation of these circRNA orthologs is no higher than that of their flanking linear exons. The previously proposed miR-7 sponge, CDR1as, is one of only two circRNAs with more miRNA sites than expected by chance, with the next best miRNA-sponge candidate deriving from a primate-specific zinc-finger gene, ZNF91. These results provide a new framework for future investigation of this intriguing topological isoform while raising doubts regarding a biological function of most circRNAs. Examination of 9 samples in 1 cell type Note: The ENCODE data we used are under GEO SuperSeries GSE26284 (all samples labeled "_cell_total"). But they were not used in the processing of the U2OS data.

2014-07-08 | E-GEOD-51584 | biostudies-arrayexpress

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