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Expression profiling of miR-294 and Mbd2 siRNA transfected Dgcr8-/- mESCs by high throughput sequencing

ABSTRACT: RNA-seq analysis on CTR, miR-294, and siMbd2 transfected Dgcr8-/- ESCs showed that Mbd2 is responsible for the change of expression of a significant portion of genes that are regulated by miR-290 cluster

ORGANISM(S): Mus musculus

PROVIDER: GSE64428 | GEO | 2015/12/31

SECONDARY ACCESSION(S): PRJNA270999

REPOSITORIES: GEO

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Expression profiling of miR-294 and Mbd2 siRNA transfected Dgcr8-/- mESCs by high throughput sequencing

Project description:RNA-seq analysis on CTR, miR-294, and siMbd2 transfected Dgcr8-/- ESCs showed that Mbd2 is responsible for the change of expression of a significant portion of genes that are regulated by miR-290 cluster mRNA profiles of CTR, miR-294 and siMbd2 transfected Dgcr8-/- ESCs were generated by deep sequencing, in duplicate, using pair-end Illumina.

2015-12-31 | E-GEOD-64428 | biostudies-arrayexpress

Single cell sequencing of Dgcr8 knockout embryonic stem cells plus/minus miR-294 or let-7

Project description:We performed Fluidigm C1 single cell sequencing analysis of wild-type and microRNA deficient (Dgcr8 knockout) mouse embryonic stem cells mock treated or transfected with either miR-294 or let-7.

2017-03-08 | GSE80168 | GEO

Let-7c and miR-294 target identification in mouse ES cells

Project description:let-7c and miR-294 were transfected into Dgcr8 -/- miRNA deficient ES cells and RNA was harvested after 12 hours the goal of this study was to identify direct and indirect targets of the let-7c and miR-294 miRNAs, we chose to harvest RNA early at 12 hours to mimize secondary effects due to ES cell differentiation, prior to performing the array experiment we found that at this time point candidate miRNA targets were maximally downregulated by qPCR

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Opposing roles of miR-290 family and muscleblind-like proteins in the regulation of embryonic stem cell-specific alternative splicing and gene expression program

Project description:RNA profiling of WT,Dgcr8-/- mESCs and miR-294 or siMbnl1/2 transfected Dgcr8-/- ESCs showed that miRNAs, especially miR-294 regulate a large number of alternative splicing events, among which muscleblind-like proteins are responsible for a significant portion. Besides, upon depletion of miRNAs, RBPs and SFs are globally downregulated in mESCs.

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Stabilizing Heterochromatin by DGCR8 Alleviates Senescence and Osteoarthritis

Project description:DiGeorge syndrome critical region 8 (DGCR8) is a critical component of the canonical microprocessor complex for microRNA biogenesis. However, the non-canonical functions of DGCR8 have not been studied. Here, we demonstrate that DGCR8 plays an important role in maintaining heterochromatin organization and attenuating aging. An N-terminal-truncated version of DGCR8 (DR8dex2) accelerated senescence in human mesenchymal stem cells (hMSCs) independent of its miRNA-processing activity, which is mediated by its C-terminal domains. Further studies revealed that DGCR8 maintained heterochromatin organization by interacting with the nuclear envelope protein Lamin B1, and heterochromatin-associated proteins, KAP1 and HP1 Overexpression of any of these proteins, including DGCR8, reversed premature senescent phenotypes in DR8dex2 hMSCs. Finally, DGCR8 was downregulated in pathologically and naturally aged hMSCs, whereas DGCR8 overexpression alleviated hMSC aging and osteoarthritis in mice. Taken together, these analyses uncovered a novel, miRNA processing-independent role for DGCR8 in maintaining heterochromatin organization and attenuating senescence. DGCR8 may therefore represent a new therapeutic target for alleviating human aging-related disorders.

2019-05-28 | PXD013856 | Pride

The effect of miR-294 and miR-181 on the transcriptome during OSK reprogramming of MEFs

Project description:Study designed to determine the immediate effects of supplementing OSK reprogramming with miR-294 or miR-181. MEFs were infected on day 0, and transfected with miR-294, miR-181 or control mimic on day 1. On day 3 RNA was extracted. OSK infected MEFs samples were compared to non-infected MEFs and to fully reprogrammed iPSCs. Performed in biological triplicate, with each sample containing different OSK viral stocks, different preparations of MEFs and independent transfections. Biological triplicate iPSCs represent three independent and clonally expanded iPSC lines. Total RNA collected with Trizol. All processing conducted at the UCLA Neuroscience Genomics Core. MouseRef-8 v2.0 Expression BeadChips.

2013-09-15 | E-GEOD-49556 | biostudies-arrayexpress

Mus musculus

Project description:Expression profiling of miR-294 and Mbd2 siRNA transfected Dgcr8-/- mESCs by high throughput sequencing

| PRJNA270999 | ENA

Biochemical Identification of Targets of Individual MicroRNAs in Mouse Embryonic Stem Cells

Project description:Mouse Embryonic Stem (ES) cells express a unique set of microRNAs (miRNAs), the miR-290-295 cluster. To elucidate the role of these miRNAs and how they integrate into the ES cell regulatory network requires identification of their direct regulatory targets. The difficulty, however, arises from the limited complementarity of metazoan miRNAs to their targets, with the interaction requiring as few as six nucleotides of the miRNA seed sequence. To identify miR-294 targets, we used Dicer1-null ES cells, which lack all endogenous mature miRNAs, and introduced just miR-294 into these ES cells. We then employed two approaches to discover miR-294 targets in mouse ES cells: transcriptome profiling using microarrays, and a biochemical approach to isolate mRNA targets associated with the Argonaute2 (Ago2) protein of the RISC (RNA Induced Silencing Complex) effector, followed by RNA-Sequencing. In the absence of Dicer1, the RISC complexes are largely devoid of mature miRNAs, and should therefore contain only transfected miR-294 and its base-paired targets. Our data suggest that miR-294 may promote pluripotency by regulating a subset of c-Myc target genes, and upregulating pluripotency-associated genes such as Lin28.

2010-02-27 | GSE20199 | GEO

Genome-wide Identification of Targets & Function of Individual MicroRNAs in Mouse Embryonic Stem Cells

2010-02-27 | GSE20048 | GEO

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