Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Reintroduction of miRNAs into Dgcr8 deficient cell lines to generate miRNA target lists

ABSTRACT: These experiments were conducted as part of a study to derive the targets of miRNAs (See also E-MTAB-418). Two independent BayGenomics mouse embryonic stem cell lines (XH157 and XG058), each bearing a gene trap between exons 9 and 10 of the Dgcr8 locus (ENSMUST00000115633, Ensembl v53) were further mutagenised by the targetted insertion of a second Dgcr8 gene trap cassette. Consequently two independent heterozygous and two independent homozygous mutant Dgcr8 cells were derived (Dgcr8tm1,gt1/+, Dgcr8tm1,gt2/+, Dgcr8gt1/tm1 and Dgcr8gt2/tm1). Subsequently individual miRNA mimics were reintroduced into the Dgcr8-depleted background. In this set of experiments, cells were allowed to recover for 10 hours. The effect of the miRNA on the mRNA expression of the cells was assessed through the comparison of the expression profile of the transfected cells to that of cells transfected with a control mimic, which had recovered over the same period. This allows the roles of individual miRNAs to be investigated.

ORGANISM(S): Mus musculus

SUBMITTER: Matthew Davis

PROVIDER: E-MTAB-968 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

SNHG9, a Papillary Thyroid Cancer Cell Exosome-Enriched lncRNA, Inhibits Cell Autophagy and Promotes Cell Apoptosis of Normal Thyroid Epithelial Cell Nthy-ori-3 Through YBOX3/P21 Pathway.

Wen Duo D Liu Wan-Lin WL Lu Zhong-Wu ZW Cao Yi-Ming YM Ji Qing-Hai QH Wei Wen-Jun WJ

Frontiers in oncology 20210504

Thyroid cancer is the most common type of endocrine malignancy. Although the general prognosis is good, the treatment of advanced disease is still challenging. Exosomes are vesicle units containing specific components that transmit information between cells. In order to explore its role in papillary thyroid cancer (PTC), our study screened exosome enriched lncRNA SNHG9 by lncRNA chip and explored its biological function. We used lncRNA chips combined with bioinformatics analysis to screen lncRNA ...[more]

PMID: 34017682

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Project description:The transition of the endothelium to a pro-inflammatory state is key to progression of chronic inflammatory diseases including rheumatoid arthritis, chronic bowel disease and atherosclerosis. In atherosclerosis it is hypothesized that low density lipoproteins (LDL) that become trapped in the intima of the blood vessels are oxidized to minimally modified LDL (mmLDL) and that these serve as an important contributing factors to endothelial dysfunction. Oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphatidylcholine (OX-PAPC), a model of the active phospholipid components of mmLDL affects the expression of hundreds of genes involved in inflammatory and other biological processes in human aortic endothelial cells (HAECs). We hypothesized that microRNAs (miRNAs) partially regulate this response. Using next generation sequencing, we identified miR-21-3p and miR-27a-5p to be induced 4-fold and 3-fold, respectively in response to OX-PAPC treatment compared to control treatment in HAECs. To identify the targets, we performed whole genome transcript profiling following transient over-expression of these two miRNAs followed by. In total, 1254 genes were down-regulated with 925 of them overlapping between the two miRNAs. Functional enrichment analysis using Gene Ontology predicted that the two miRNAs were involved in the regulation of NF-κB signaling. We characterized the Toll/interleukin-1 receptor (TIR) domain-containing adaptor protein TICAM2 as a direct target of miR-21-3p and miR-27a-5p. Furthermore, we showed that over-expression of miR-21-3p and miR-27a-5p lead to decreased p65 translocation to the nucleus and decreased the expression of known NF-κB downstream target genes confirming both miRNAs’ role in negatively regulating NF-κB signaling in endothelial cells. mRNA expression profiling of human aortic endothelial cells from two separate donors that were transfected with 1 nM microRNA mimics and negative control. The miRIDIAN mimics used were miR-21-3p (Catalog Number:C-301023-01-0005), miR-27a-5p (Catalog No: C-301028-01-0005), negative control (Catalog No: CN-001000-01-05)

Dataset Information

Reintroduction of miRNAs into Dgcr8 deficient cell lines to generate miRNA target lists

Publications

SNHG9, a Papillary Thyroid Cancer Cell Exosome-Enriched lncRNA, Inhibits Cell Autophagy and Promotes Cell Apoptosis of Normal Thyroid Epithelial Cell Nthy-ori-3 Through YBOX3/P21 Pathway.

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