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RNA immunoprecipitation sequencing (RIP seq) using anti-Argonaute2 (Ago2) antibody to identify the potential targets of kshv-miR-K12-1-5p in AC16 cells

ABSTRACT: RNA immunoprecipitation sequencing (RIP seq) using anti-Argonaute2 (Ago2) antibody to identify the potential targets of kshv-miR-K12-1-5p in AC16 cells

PROVIDER: PRJNA828132 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR18811428_1.fastq.gz	Fastqsanger.gz
	SRR18811428_2.fastq.gz	Fastqsanger.gz
	SRR18811429_1.fastq.gz	Fastqsanger.gz
	SRR18811429_2.fastq.gz	Fastqsanger.gz
	SRR18811430_1.fastq.gz	Fastqsanger.gz

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RNA immunoprecipitation sequencing (RIP seq) using anti-Argonaute2 (Ago2) antibody to identify the potential targets of kshv-miR-K12-1-5p in AC16 cells

Project description:RNA immunoprecipitation sequencing (RIP seq) using anti-Argonaute2 (Ago2) antibody to identify the potential targets of kshv-miR-K12-1-5p in AC16 cells.

2022-04-22 | GSE201041 | GEO

Argonaute2 (Ago2) immunoprecipitation (IP) assay using miR-130b overexpressing A549 cells.

Project description:Argonaute2 (Ago2) immunoprecipitation (IP) assay using miR-130b overexpressing A549 cells.

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Argonaute2 attenuates active transcription by limiting RNA Polymerase II elongation in Drosophila melanogaster [ChRIPseq-AGO2]

Project description:Increasing lines of evidence support that Argonaute2 (AGO2) harbors several nuclear functions in metazoa. In particular, Drosophila AGO2 modulates transcription of developmentally regulated genes; however, the molecular mechanisms behind AGO2 recruitment into chromatin and its function in transcription have not been deeply explored. In this study, we show that Drosophila AGO2 chromatin association depends on active transcription. In order to gain insight into how AGO2 controls transcription, we performed differential ChIP-seq analysis for RNA Polymerase II (Pol II) upon depletion of AGO2. Interestingly, we find specific accumulation of the elongating but not initiating form of Pol II after AGO2 knockdown, suggesting that AGO2 impairs transcription elongation. Finally, AGO2 also affects Negative Elongation Factor (NELF) chromatin association but not the Cyclin Dependent Kinase 9 (CDK9). Altogether, these results provide key insights into the molecular role of AGO2 in attenuating elongation of certain actively transcribed genes.

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Expression data from Dicer and Ago2-knockdown HeLa cells

Project description:Dicer and Argonaute2 (Ago2) gene is involving in microRNA (miRNA) maturation. Knockdown of these genes has great impact on miRNA expression profiles. We used microarrays to detail the miRNA expression profiles in Dicer- and Ago2-knockdown HeLa cells and demonstarted that the significant difference between Ago2-knockdown and Dicer- and Ago2-co-knockdown HeLa cells were not found.

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Essential role for Argonaute2 protein in mouse oogenesis

Project description:Argonaute2 proteins (Ago2) are key component of RNA-induced gene silencing complexes (RISCs), which is crucial for microRNAs to repress target genes. The function of Ago2 for oogenesis is unknown due to early embryonic lethal of Ago2 knockout mice. Here we show the effect of loss of Ago2 in mouse oogenesis by specific deletion of it in growing oocyte. Although the Ago2-deficient oocytes can develop to mature oocytes, they have abnormal spindle and the chromosomes that can’t cluster together properly, which are very similar to the phenotype of Dicer-deficient oocytes. We checked miRNA expression profile and found that in Ago2-deficient oocyte, the expression of most miRNAs reduced more than 80%. To understand the downstream genes regulated by Ago2, we used microarray on Ago2-deficient oocyte and found that 512 genes were upregulated and 1073 genes were downregulated (FC>2, P<0.05). Our data proved that Ago2 have a key function for mouse oogenesis through globally regulating miRNA stability and function.

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RNAi-independent role for Argonaute2 in CTCF/CP190 chromatin insulator function

Project description:Chromatin insulators and Polycomb group (PcG) complexes control nuclear organization to effect changes in gene expression. In Drosophila, RNA silencing pathways influence long range interactions mediated by PcG proteins and nuclear localization of the gypsy insulator; however, the underlying mechanisms are unknown. Here, we identify a singular requirement for Argonaute2 (AGO2) for the activity of the CCCTC-binding factor (CTCF)/Centrosomal protein 190 (CP190) dependent Fab-8 insulator. AGO2 and CP190 interact physically, and genome wide localization of AGO2 by chromatin immunoprecipitation and sequencing (ChIP-seq) reveals extensive colocalization of AGO2 with insulators and Polycomb Response Elements (PREs) but minimal overlap with regions of endogenous small interfering RNA (endo-siRNA) production. Finally, depletion of either CTCF or CP190 results in loss of AGO2 association with insulators, PREs, and other cis-regulatory regions. Our findings suggest that Dicer-independent recruitment of AGO2 to chromatin by insulator proteins promotes the definition of transcriptional domains throughout the genome. ChIP-seq of AGO2 in two Drosophila cell types (S2 and S3)

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Project description:Here we identify a Dicer-independent miRNA biogenesis pathway that employs the slicer catalytic activity of Argonaute2 (Ago2). To uncover Dicer-independent miRNAs, we sequenced small RNAs in wild type, maternal-zygotic dicer (MZdicer) and MZago2 mutants, using zebrafish as a model system. We find that, in contrast to other miRNAs, miR-451 levels were increased in MZdicer but drastically reduced in the MZago2 mutants. We show that pre-miR-451 processing requires Ago2 catalytic activity in vivo. MZago2 mutant embryos display delayed erythrocyte maturation that can be rescued by wild type Ago2 or miR-451 duplex but not catalytically dead Ago2. We propose that Ago2-mediated cleavage of a subset of pre-miRNAs, followed by uridylation and trimming, generates functional miRNAs in a Dicer-independent manner. Examination of small RNAs (18 to 35 nucleotides) in 3 different zebrafish genotypes (wild type, MZago2, MZdicer) at 48 hours post-fertilization.

2010-07-01 | E-GEOD-21503 | biostudies-arrayexpress

Argonaute2 and LaminB modulate gene expression by controlling chromatin topology (ChIP-seq)

Project description:Drosophila Argonaute2 (AGO2) is known to regulate expression of certain loci in an RNA interference-independent manner, but its genome-wide function on chromatin remains unknown. Here, we identified RNA Polymerase II (Pol II) and LaminB as AGO2 nuclear interactors. AGO2 and LaminB attenuate transcription at borders between both LaminB-associated domains (LADs) and topologically-associated domains (TADs). We identified a somatic target of AGO2 transcriptional repression, nht, which is immersed in a LAD located within a repressive TAD. Null mutation of AGO2 leads to ectopic expression of spermatogenesis genes dependent on nht. Finally, depletion of either AGO2 or LaminB results in reduced looping interactions within the TAD as well as ectopic inter-TAD interactions. Overall, our findings reveal coordination of AGO2 and LaminB to dictate overall genome architecture and thereby regulate gene expression.

2018-02-27 | GSE101365 | GEO

RNA sequencing of AC16 and AC16-CAR cells

Project description:RNA sequencing of AC16 and AC16-CAR cells

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A novel miRNA processing pathway independent of Dicer requires Argonaute2 catalytic activity

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