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The network of genes regulated by Ago1 in metastatic prostate cancer PC3

ABSTRACT: The objective of this study was to evaluate the consequences of Ago1 knockdown in metastatic prostate cancer cells PC3. To this end we profiled RNA from PC3 cells 72 hours following the transfection with a scramble siRNA (siGL3) and a siRNA targeting Ago1 (siAgo1).

ORGANISM(S): Homo sapiens

PROVIDER: GSE57885 | GEO | 2015/05/22

SECONDARY ACCESSION(S): PRJNA248343

REPOSITORIES: GEO

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The network of genes regulated by Ago1 in metastatic prostate cancer PC3

Project description:The objective of this study was to evaluate the consequences of Ago1 knockdown in metastatic prostate cancer cells PC3. To this end we profiled RNA from PC3 cells 72 hours following the transfection with a scramble siRNA (siGL3) and a siRNA targeting Ago1 (siAgo1). We profiled RNA from PC3 cells 72 hours following the transfection with a scramble siRNA (siGL3) and a siRNA targeting Ago1 (siAgo1). siAgo1 vs siGL3 gene expression profiling, two technical replicates with dye swap labeling scheme.

2015-05-22 | E-GEOD-57885 | biostudies-arrayexpress

Chromatin accessiblity of 3T3-L1 cells with Kdm5c knockdown

Project description:One day prior to confluency, non-targeting (scramble control) siRNA or Kdm5c siRNA were transfected into 3T3-L1 cells and chromatin structure was assessed by ATAC-seq at 24hr, 48hr, and 72 hr post transfection.

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Project description:Patient-derived glioma stem-like cell (GSC-11), a kind gift of Dr. Lang at UT MD Anderson Cancer Center, was subjected to a transient transfection to down-regulate SOX2 expression. Specific human SOX2 siRNA and a non-targeting control siRNA (si-Scramble) were used in four independent experiments. The cells were then cultured for 72 h after transfection and subjected to the miRNA array analysis.

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MELK and UBE2C gene knockdown in prostate cancer cell line PC3

Project description:In order to address the putative role of MELK and UBE2C in prostate cancer development and progression, we performed functional analysis upon siRNA-based knockdown, and searched for downstream genes and processes by microarray experiments. RNAi-based inhibition of MELK and UBE2C was efficient in PC3 prostate cancer cells and decreased transcriptional level down to about 30% remaining expression level. Illumina microarray experiments were done upon siRNA based knockdown 48h after transfection of PC3 cells in triplicates.

2012-10-19 | E-GEOD-32873 | biostudies-arrayexpress

Total and AGO1 immunoprecipitated small RNA from Arabidopsis thaliana wild-type and ago1-57 mutant

Project description:In Arabidopsis thaliana, ARGONAUTE1 (AGO1) plays a central role[AQ1] in microRNA (miRNA) and small interfering RNA (siRNA)- mediated silencing and is a key component in antiviral responses. The polerovirus F-box P0 protein triggers AGO1 degradation as a viral counterdefense. Here, we identified a motif in AGO1 that is required for its interaction with the S phase kinase-associated protein1-cullin 1-F-box protein (SCF) P0 (SCFP0) complex and subsequent degradation. The AGO1 P0 degron is conserved and confers P0-mediated degradation to other AGO[AQ2] proteins. Interestingly, the degron motif is localized in the DUF1785 domain of AGO1, in which a single point mutation (ago1-57, obtained by forward genetic screening) compromises recognition by SCFP0. Recapitulating formation of the RNA-induced silencing complex in a cell-free system revealed that this mutation impairs RNA unwinding, leading to stalled forms of AGO1 still bound to double-stranded RNAs. In vivo, the DUF1785 is required for unwinding perfectly matched siRNA duplexes, but is mostly dispensable for unwinding imperfectly matched miRNA duplexes. Consequently, its mutation nearly abolishes phased siRNA production and sense transgene posttranscriptional gene silencing. Overall, our work sheds new light on the mode of AGO1 recognition by P0 and the in vivo function of DUF1785 in RNA silencing.

2018-06-22 | GSE104015 | GEO

Arabidopsis AGO1 N-terminal Poly-Q domain promotes phase separation and association with stress granules during heat stress

Project description:In Arabidopsis thaliana, ARGONAUTE1 (AGO1) plays a central role in microRNA (miRNA) and small interfering RNA (siRNA)-mediated silencing. Nuclear AGO1 is loaded with miRNAs and exported to the cytosol where it associates to the rough ER to conduct miRNA-mediated translational repression, mRNA cleavage and biogenesis of phased siRNAs. These latter, as well as other cytosolic siRNAs, are loaded into cytosolic AGO1, but in which compartment this happens is not known. Moreover, the effect of stress on AGO1 localization is still unclear. Here, we show that heat stress (HS) promotes AGO1 protein accumulation, which co-localize with components of the siRNA bodies and of stress granules (SGs). AGO1 does not need SGS3, a key component of siRNA bodies, to efficiently form condensates during HS. Instead, we found that the still poorly characterized N-terminal Poly-Q domain of AGO1, which contains a prion-like domain, is sufficient to undergo phase separation. Moreover, an exposure of 1 hour to HS only moderately affected AGO1 loading by miRNAs and target cleavage, suggesting that its localization in condensates protects AGO1 rather than promote its activity in reprograming gene expressing during stress. Collectively, our work shed new light on the impact of high temperature on a main effector of RNA silencing in plants.

2024-06-22 | PXD048594 | Pride

Pimary human keratinocyte knockdown with deltaNp63 specific siRNA

Project description:Human ΔNp63-specific siRNA was obtained from Invitrogen (Carlsbad, CA; sense: 5′-ACAAUGCCCAGACUCAAUU-3′; antisense: 5′-AAUUGAGUCUGGGCAUUGU-3′). Scrambled sequence siRNA for the negative control was purchased from Invitrogen. Transfections were performed using Lipofectamine RNAiMAX (Invitrogen) in Opti-MEM (GIBCO) at 40 nmol/L according to the manufacturer's instructions. The culture medium was replaced at 6 h after siRNA transfection. mRNA was extracted from NHEKs transfected with ΔNp63-specific or scramble sequence siRNA at 72 h after transfection. Microarray slides were scanned using a 3D-GENE human 25k (TORAY, Tokyo, Japan) and microarray images were automatically analyzed with AROSTM, version 4.0 (Operon Biotechnologies, Tokyo, Japan).

2020-12-08 | GSE162900 | GEO

Gene expression profiling in cells with Ago1 knockdown (Affymetrix)

Project description:Argonautes, a family of highly evolutionarily conserved proteins, are the central platform for small RNA-mediated gene regulatory mechanisms which occur mainly in the cytoplasm. To understand a potential role of Argonaute 1 (Ago1) protein in the nucleus of mammalian cells in regulating gene transcription and epigenetics, we performed integrated analyses of Ago1 ChIP-sequencing (GSE40536) and gene expression profiling in cells depleted of Ago1. For gene expression profiling, we knocked down the expression of Ago1 by siRNA in PC-3 cells and compared gene expression profiles in the cells depleted of Ago1 and cells receiving control treatments. We found that Ago1 depletion resulted in more downregulated genes which were enriched in gene responsible for promoting cell cycle progression, DNA replication and repair, and response to endogenous stimuli. Integrated analyses of Ago1-chromosomal interactions and gene expression changes in response to Ago1 depletion reveal a significant correlation between Ago1-bound genes and genes altered by Ago1 depletion. These genes are significantly enriched in cancer-related pathways. Our data suggests a nuclear function of Ago1 in regulating gene transcription. PC-3 cells were mock transfected or transfected with an Ago1 specific siRNA or a control siRNA. Each treatment was done in duplicates.

2013-09-27 | E-GEOD-42600 | biostudies-arrayexpress

miR-145 transfected PC3 cells

Project description:It is aimed to reveal overall trancriptional change in prostate cancer PC3 cells after ectopic expression of miR-145 Pre-mir-145 transfected PC3 cells were collected at 8, 16 and 24 hours after transfection and control untrasfected PC3 cells were used.

2014-06-15 | E-GEOD-58295 | biostudies-arrayexpress

Degradome sequencing of Arabidopsis ago1-27 mutant

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