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Genome-wide analysis of miRNA-mRNA interactions in the marrow microenvironment

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Homo sapiens

PROVIDER: GSE48249 | GEO | 2013/09/01

SECONDARY ACCESSION(S): PRJNA209350

REPOSITORIES: GEO

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Genome-wide analysis of miRNA-mRNA interactions in the marrow microenvironment

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2013-09-01 | E-GEOD-48249 | biostudies-arrayexpress

Genome-wide analysis of miRNA-mRNA interactions in marrow stromal cells.

Project description:Regulation of hematopoietic stem cell proliferation, lineage commitment, and differentiation in adult vertebrates requires extrinsic signals provided by cells in the marrow microenvironment (ME) located within the bone marrow. Both secreted and cell-surface bound factors critical to this regulation have been identified, yet control of their expression by cells within the ME has not been addressed. Herein we hypothesize that microRNAs (miRNAs) contribute to their controlled expression. MiRNAs are small noncoding RNAs that bind to target mRNAs and downregulate gene expression by either initiating mRNA degradation or preventing peptide translation. Testing the role of miRNAs in downregulating gene expression has been difficult since conventional techniques used to define miRNA-mRNA interactions are indirect and have high false-positive and negative rates. In this report, a genome-wide biochemical technique (high-throughput sequencing of RNA isolated by cross-linking immunoprecipitation or HITS-CLIP) was used to generate unbiased genome-wide maps of miRNA-mRNA interactions in two critical cellular components of the marrow ME: marrow stromal cells and bone marrow endothelial cells. Analysis of these datasets identified miRNAs as direct regulators of JAG1, WNT5A, MMP2, and VEGFA; four factors that are important to ME function. Our results show the feasibility and utility of unbiased genome-wide biochemical techniques in dissecting the role of miRNAs in regulation of complex tissues such as the marrow ME.

| S-EPMC4127404 | biostudies-literature

Genome-wide analysis of miRNA-mRNA interactions in the marrow microenvironment (HTS)

Project description:We report genome-wide analysis of miRNA-mRNA interactions in the hematopoietic marrow microenvironment (ME) by employing the biochemical technique. High-throughput sequencing of RNA isolated by crosslinking immunoprecipitation or HITS-CLIP. Specifically, we analyzed 3 kinds of stromal cells (two human stromal cell lines named HS5 and HS27a and primary mesenchymal stromal cells (MSC) from normal donors) and two types of endothelial cells (TrBMEC, a human bone marrow endothelial cell line) and HUVEC (Human Umbilical Vein Endothelial Cells). Immune precipitation (IP) of Argonaut proteins (AGO1-4) was performed by anti-AGO monoclonal antibody 2A8. IP was performed on cells cross-linked by short-wave ultraviolet radiation (UV) 365 nM. Protocol was modified from as originally published by Chi et al Nature. 2009 Jul 23;460(7254):479-86.

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Genome-Wide Profiling of miRNA-mRNA regulatory network in Skeletal Muscle with Aging

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