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MRNAs associated with Ago2: Ago2, Ago2+miR-1, Ago2+miR-124, mock transfected

ABSTRACT: As a core RISC component, Ago2 associates with miRNAs and target mRNAs. To identify these mRNAs, we ran lysate from HEK293T cells over a FLAG resin from 2 conditions: +FLAG-Ago2, +mock transfection. To identify mRNAs associated with specific miRNAs, we ran lysate from HEK293T cells over a FLAG resin from 2 conditions: +FLAG-Ago2 & miR-1, and +FLAG-Ago2 & miR-124. Set of arrays that are part of repeated experiments Compound Based Treatment: mock transfected Keywords: Biological Replicate

ORGANISM(S): Homo sapiens

PROVIDER: GSE11079 | GEO | 2008/04/08

SECONDARY ACCESSION(S): PRJNA108827

REPOSITORIES: GEO

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mRNAs associated with Ago2: Ago2, Ago2+miR-1, Ago2+miR-124, mock transfected

Project description:As a core RISC component, Ago2 associates with miRNAs and target mRNAs. To identify these mRNAs, we ran lysate from HEK293T cells over a FLAG resin from 2 conditions: +FLAG-Ago2, +mock transfection. To identify mRNAs associated with specific miRNAs, we ran lysate from HEK293T cells over a FLAG resin from 2 conditions: +FLAG-Ago2 & miR-1, and +FLAG-Ago2 & miR-124. Set of arrays that are part of repeated experiments Compound Based Treatment: mock transfected Keywords: Biological Replicate Biological Replicate Computed

2008-04-08 | E-GEOD-11079 | biostudies-arrayexpress

Expression data HEK293T cells transfected with either Ago2, Ago2 + miR-1, Ago2 + miR-124, mock

Project description:HEK293T cell expression data from four conditions: mock transfection, Ago2 transfection, Ago2 & miR-1 transfection, and Ago2 & miR-124 transfection. RNA was isolated directly from 10cm dishes using Trizol reagent and amplified. Set of arrays that are part of repeated experiments Compound Based Treatment: FLAG-Ago2 transfection Keywords: Biological Replicate

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Expression data HEK293T cells transfected with either Ago2, Ago2 + miR-1, Ago2 + miR-124, mock

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miRNA arrays: Ago2 transfected vs. Mock, Ago2 IP vs. total RNA

Project description:miRNAs were enriched from HEK293T cells using the ambion FLASHPAGE fractionater after mock transfection, Ago2 transfection, and FLAG-Ago2 IP. miRNAs were labeled and hybridized: Ago2 transfected vs. mock transfection and Ago2 transfected vs. Ago2 IPs. Set of arrays that are part of repeated experiments Keywords: Biological Replicate

2008-04-08 | GSE11081 | GEO

miRNA arrays: Ago2 transfected vs. Mock, Ago2 IP vs. total RNA

2008-04-07 | E-GEOD-11081 | biostudies-arrayexpress

Identification of miRNA targets in breast cancer cells (Ago2-IP)

Project description:miRNAs regulate mRNA stability and translation through the action of the RNAi-induced silencing complex. In this study, we systematically identified endogenous miRNA target genes by using AGO2 immunoprecipitation (AGO2-IP) and microarray analyses in two breast cancer cell lines, MCF7 and MDA-MB-231, representing luminal and basal-like breast cancer, respectively. The expression levels of ~70% of the AGO2-IP mRNAs were increased by DROSHA or DICER1 knockdown. In addition, integrated analysis of miRNA expression profiles, mRNA-AGO2 interaction, and the 3'-UTR of mRNAs revealed that >60% of the AGO2-IP mRNAs were putative targets of the fifty most abundantly expressed miRNAs. To identify mRNAs associated with AGO2, cell lysate was precleaned with control IgG and immunoprecipitated with anti-human Ago2 (Clone 2E12-1C9, Abnova). Total RNA from cell lysate or coimmunoprecipitated with AGO2 was extracted with Trizol and subjected to microarray analysis, with three biological repeats for each experimental condition.

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Identification of mRNAs Recruited to Argonaute 2 by Specific microRNAs and Corresponding Changes in Transcript Abundance

Project description:This SuperSeries is composed of the following subset Series: GSE11079: mRNAs associated with Ago2: Ago2, Ago2+miR-1, Ago2+miR-124, mock transfected GSE11080: Expression data HEK293T cells transfected with either Ago2, Ago2 + miR-1, Ago2 + miR-124, mock GSE11081: miRNA arrays: Ago2 transfected vs. Mock, Ago2 IP vs. total RNA Keywords: SuperSeries Refer to individual Series

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microRNA binding by wildtype Ago2 and mutant Ago2 in human cells

Project description:Ago2 binds mature microRNAs (miRNAs) to regulate gene expression. A mutant Ago2 is constructed to render it unable to bind miRNAs and inhibit mRNA translation. Human 293T cells were transiently transfected with a plasmid that encoded a FLAG-tagged wildtype human Ago2 or mutant Ago2. FLAG IP was preformed, and assoicated RNA was isolated and subjected miRNA microarray analysis.

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A nuclear role for the Argonaute protein AGO2 in mammalian gametogenesis

Project description:The reduced sperm count observed in Ago2 cKO mice implies that AGO2 has non-redundant functions in the male germ line. Because AGO2 is a key protein in the RNA interference (RNAi) pathway, we first postulated that AGO2 loss disrupts normal transcriptional and translational dynamics of target mRNAs relevant to sperm maturation. To address this hypothesis, we took advantage of the apparently normal spermatogenesis in Ago2 cKO mice, which allowed us to purify matched meiotic and post-meiotic germ cells from Ago2 cKO and wild type controls. We examined changes in the transcriptome and proteome of these two spermatogenic stages in Ago2 cKO relative to control mice using RNA-seq and quantitative mass spectrometry (MS). To further examine if the changes in mRNA and protein levels detected in Ago2 cKO germ cells was due to a loss of regulation by the canonical AGO2-miRNA pathway, we characterized AGO2 protein interactors by AGO2 immunoprecipitation-mass spectrometry (IP-MS) in cytoplasmic and nuclear fractions of post-meiotic cells

2022-08-01 | PXD027935 | Pride

Ago2 associated gene transcripts in two Hodgkin lymphoma cell lines

Project description:The aim of the study is to identify miRNA targets in Hodgkin lymphoma cell lines. By immunoprecipitation of wild type Ago2, it is expected to pull down the Ago2 associated gene transcripts. Through microarray analysis, the Ago2 associated gene transcripts identified are expected to be miRNA targets. Keywords: Ribonucleoprotein Immunoprecipitation - Gene Chip (RIP-Chip) RNA isolated from the Ago2 immunoprecipitated (IP) fraction is hybridized against the RNA from total cell lysate (T) fraction. The experiment is performed in two independent Hodgkin lymphoma cell lines, L1236 and L428. In addition, RNA of flow through (FT) fraction of L1236 is hybridized against RNA of total cell lysate (T) fraction of L1236 to demonstrate the specificity/enrichment seen in the IP fraction and not in the FT fraction. All hybridizations is done with a dye swap design.

2010-04-08 | E-GEOD-14409 | biostudies-arrayexpress

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