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SmRNAs in wildtype and RDR6-15 knockout Arabidopsis thaliana Col-0 leaf

ABSTRACT: This set consists of small RNAs sequenced from two replicates of wildtype and two replicates of RDR6-15 knockout Arabidopsis thaliana Col-0 leaf samples. RDR6 is required for the production of tasRNAs (trans-acting small RNAS) and so tags associated with the tasRNA loci should be severely down-regulated or absent in the knockout compared to wildtype. The set can thus be used as containing known true positives for testing differential expression detection methods. Examination of smRNA in 2 replicates wildtype and 2 replicates RDR6-15 knockout

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Thomas Hardcastle

PROVIDER: E-GEOD-16959 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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baySeq: empirical Bayesian methods for identifying differential expression in sequence count data.

Hardcastle Thomas J TJ Kelly Krystyna A KA

BMC bioinformatics 20100810

<h4>Background</h4>High throughput sequencing has become an important technology for studying expression levels in many types of genomic, and particularly transcriptomic, data. One key way of analysing such data is to look for elements of the data which display particular patterns of differential expression in order to take these forward for further analysis and validation.<h4>Results</h4>We propose a framework for defining patterns of differential expression and develop a novel algorithm, baySe ...[more]

PMID: 20698981

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smRNAs in wildtype and RDR6-15 knockout Arabidopsis thaliana Col-0 leaf

Project description:This set consists of small RNAs sequenced from two replicates of wildtype and two replicates of RDR6-15 knockout Arabidopsis thaliana Col-0 leaf samples. RDR6 is required for the production of tasRNAs (trans-acting small RNAS) and so tags associated with the tasRNA loci should be severely down-regulated or absent in the knockout compared to wildtype. The set can thus be used as containing known true positives for testing differential expression detection methods.

2010-11-22 | GSE16959 | GEO

Gene expression of flower buds between rdr6-15 and wild-type under drought stress condition

2014-09-02 | GSE57174 | GEO

Arabidopsis thaliana

Project description:smRNAs in wildtype and RDR6-15 knockout Arabidopsis thaliana Col-0 leaf

| PRJNA117297 | ENA

Gene expression of flower buds between rdr6-15 and wild-type under drought stress condition

Project description:To understand ta-siRNA-mediated regulation network in response to abiotic stress during flower development, gene expression profiles of flowering buds between rdr6-15 and wild-type under moderate drought stress and control condition were measured using microarray. 512 genes were bilaterally regulated by drought stress signal and RDR6-mediated signal. rdr6-15 and wild-type plants (Arabidopsis thaliana ecotype Columbia) were grown on 30g of vermiculite soil of plastic pot for 2 weeks. Then, water depletion (drought stress treatment) was applied for 1 week. Water content of the soil was decreased to less than 20%. 10 flower buds were collected for each experiment. The experiment was repeated 3-times.

2014-09-02 | E-GEOD-57174 | biostudies-arrayexpress

Arabidopsis small RNAs associated with RDR6-dependent RNA-directed DNA Methylation

Project description:Background: The majority of plant transposable elements (TEs) are found in a silenced state that is epigenetically propagated by the maintenance of symmetrical DNA methylation. TE methylation is established and reinforced by a mechanism of RNA-dependent DNA methylation, which is dependent on transcription of the PolIV RNA polymerase. Recently, a pathway has been described that initiates de novo DNA methylation dependent on components of the RNAi post-transcriptional silencing pathway, independent of PolIV. To define the function of this new pathway, we have focused on the RDR6 protein, which plays a central role in the pathway we refer to as RDR6-dependent RNA-directed DNA Methylation (RDR6-RdDM). Methods: We have sequenced total small RNAs from wild-type and three mutant genotypes: ddm1, ddm1/rdr6, and rdr6. Conclusions: We demonstrate that RDR6-RdDM utilizes 21 and 22 nucleotide siRNAs to de novo methylate actively transcribing TEs. In addition, we find that the RDR6-RdDM pathway functions in the efficient initiation and re-establishment of TE transcriptional silencing, while it is not necessary to maintain trans-generational epigenetic silencing.

2013-04-11 | GSE41755 | GEO

Principles Governing Dicer-2 Substrate Selection are Revealed by Characterizing the RNAi Response to Virus Infection and Exogenous dsRNA

Project description:Background: RNA silencing pathways play critical roles in gene regulation, virus infection, and transposon control. RNA interference (RNAi) is mediated by small interfering RNAs (siRNAs), which are liberated from double stranded (ds) RNA precursors by Dicer and direct the RNA-induced silencing complex (RISC) to target transcripts. Recent efforts have uncovered important principles governing small RNA (smRNA) sorting into RISC, yet mechanisms defining substrate selection by Dicer proteins remain uncharacterized. Methodology: To better characterize Dicer-2 substrates in Drosophila, we examined the antiviral RNAi response, which generates virus-derived siRNAs from viral RNA. Using high-throughput sequencing, we found that diverse viruses were uniquely targeted; substrates included dsRNA replication intermediates and intramolecular RNA stem loops. smRNA distribution patterns from viral and synthetic dsRNA precursors were highly reproducible, and machine learning techniques identified characteristics of precursor molecules and smRNA duplexes important in determining relative smRNA abundance. Significance: To our knowledge, this study provides the first description of the rules governing Dicer-2 substrate selection, which has important implications for exogenous RNA silencing technologies and the development of smRNA-based antiviral therapeutics. virus-derived siRNA (vsiRNA) expression comparison between control and 4 different virus-infected cells in control as well as 5 different RNAi pathway protein knock-downs in Drosophila dl1 cells

2013-01-31 | E-GEOD-28957 | biostudies-arrayexpress

smRNA under high osmotic stress

Project description:To identifiy osmotic stress responsive smRNAs, we used a deep-sequencing technique to profile small RNA populations in leaf and root tissues of plants under high osmotic stress and control conditions. We treated 30day-old plants with high osmotic stress and sampled leaves and roots from the same plant with 3 biologial replicates. Then, 3 replicates were pooled and total RNA was extracted and prepared for smRNA deep sequencing. After normalization and annotation, we selected potential osmotic stress responsive smRNAs.

2012-09-19 | E-GEOD-36560 | biostudies-arrayexpress

Genome-wide double-stranded RNA sequencing reveals the functional significance of base-paired RNAs in Arabidopsis

Project description:The functional structure of all biologically active molecules is dependent on intra- and inter-molecular interactions. This is especially evident for RNA molecules whose functionality, maturation, and regulation requires formation of correct secondary structure through encoded base-pairing interactions. Unfortunately, intra- and inter-molecular base-pairing information is lacking for most RNAs. Here, we use high-throughput sequencing to interrogate all base-paired RNA in Arabidopsis thaliana, and identify ~200 new small (sm)RNA-producing substrates of RNA-DEPENDENT RNA POLYMERASE 6. Our comprehensive analysis of paired RNAs reveals conserved functionality within introns and both 5’ and 3’ untranslated regions (UTRs) of mRNAs, as well as a novel population of functional RNAs, many of which are the precursors of smRNAs. Finally, we identify intra-molecular base-pairing interactions to produce a genome-wide collection of RNA secondary structure models. These findings highlight the importance of base-paired RNAs in eukaryotes, and present an approach that should be widely applicable for the analysis of this key structural feature of RNA. Double-stranded (dsRNA) specific RNA sequencing (dsRNA-seq) in the unopened flowerbuds of wild-type col-0 plants and rdr6 mutant plants, including 2 samples of col-0 dsRNA (1X- or 2X- Ribominus-treated samples) and 1 sample of rdr6 dsRNA. Corresponding two smRNA libraries (smRNA-seq) of both wild-type col-0 plants and rdr6 mutant plants of the same tissue are also presented.

2010-10-04 | E-GEOD-23439 | biostudies-arrayexpress

Transcriptome analysis of two mutants rdr6/sgs2 and sgs3 impaired in post-transcriptional gene silencing (PTGS)

Project description:Identification of new genes regulated by RDR6 and SGS3 (two genes involved in PTGS) by analysis of the transcriptome of rdr6-1 and sgs3-1 mutants compared to wild-type plants in different tissues (flower and leaves). The comparison between transcriptome of rdr6-1 and sgs3-1 mutant alleles impaired in PTGS and development (juvenile-to-adult transition) and transcriptome of rdr6-5 and sgs3-3 alleles impaired only in PTGS would allowed identification of genes involved in the developmental default (zip phenotype) of the null alleles (rdr6-1 and sgs3-1 mutants). - Transcriptome of rdr6 and sgs3 mutants will be compared to the transcriptome of wild-type plants in flower and leaves. Further-more the comparison between transcriptome of rdr6-1 and sgs3-1 mutant alleles impaired in PTGS and development (juvenile-to-adult transition) and transcriptome of rdr6-5 and sgs3-3 alleles impaired only in PTGS will be done for flowers and leaves. Keywords: gene knock in (transgenic)

2007-05-31 | GSE7936 | GEO

Virus infection triggers widespread silencing of host genes by a distinct class of endogenous siRNAs in Arabidopsis

Project description:Virus infection triggers widespread silencing of host genes by a distinct class of endogenous siRNAs in Arabidopsis This study constructed and sequenced two independent small RNA libraries from the upper uninoculated leaves of (i) WT Arabidopsis plants 14 d after mock inoculation and of (ii) rdr1 rdr6 plants 14 d after infection with Fny CMV-Δ2b, and one library each from the upper uninoculated leaves of (iii) WT, (iv) rdr1, and (v) rdr6 plants 14 d after infection with TuMV-GFP. Coimmunoprecipitation with FLAG- and HA-specific antibodies was used to obtain (vi,vii) AGO1 and (viii,ix) AGO2 complexes, respectively, from the FLAG-AGO1/ago1-36 and HA-AGO2 plants 14 d after infection with Fny CMV-Δ2b for extracting total loaded small RNAs for the construction and sequencing of small RNA libraries.

2014-08-30 | E-GEOD-60929 | biostudies-arrayexpress

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