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PiRNAs and transgenerational RNA interference protect essential genes from RNA silencing in C. elegans

ABSTRACT: This data series contains small RNA high-throughput sequencing data derived from cell lysates and FLAG::HRDE-1 immunoprecipitates (IP) after resetting RNAi in the presence or absence of piRNAs in C. elegans.

ORGANISM(S): Caenorhabditis elegans

PROVIDER: GSE63114 | GEO | 2015/06/30

SECONDARY ACCESSION(S): PRJNA266654

REPOSITORIES: GEO

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piRNAs and transgenerational RNA interference protect essential genes from RNA silencing in C. elegans

Project description:This data series contains small RNA high-throughput sequencing data derived from cell lysates and FLAG::HRDE-1 immunoprecipitates (IP) after resetting RNAi in the presence or absence of piRNAs in C. elegans. Small RNAs were isolated from adult wild type and mutant or transgenic C. elegans and subjected to Illumina HiSeq sequencing. The series contains fastq and tab-separated files for 10 libraries.

2015-06-30 | E-GEOD-63114 | biostudies-arrayexpress

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P granules protect RNA interference genes from silencing by piRNAs

Project description:This SuperSeries is composed of the SubSeries listed below.

2019-07-23 | GSE134638 | GEO

RNA immunoprecipitation to identify RHA-binding transcripts in HEK293 cells

Project description:Given that RHA regulates translation by binding a PCE located at the 5’ UTR of the target transcripts a genome-wide screen was performed to identify mRNAs that bind RHA in vivo. The results from four experiments with samples obtained in four independent RNA immunoprecipitations identified 375 transcripts that co-immunoprecipitate with FLAG RHA. Since N-terminal FLAG tagged RHA specifically co-immunoprecipitates PCE-containing mRNAs HEK293 cells were transfected with a CMV-FLAG-RHA construct. Cytoplasmic lysates were immunoprecipitated with anti-FLAG beads. RNA was extracted from the immunoprecipitate and used to probe a human Agilent expression arrays. An immunoprecipitation with cells transfected with empty FLAG plasmid was used as negative control.

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Project description:To explore the roles of piRNAs in regulating 22G-RNA production and transgenerational gene silencing, we used RNA-seq to assess changes in small RNA and mRNA levels in piwi/prg-1 and mutator mutants at an early generation and again at a late generation. We identified that in the absence of piRNAs, many genes gain 22G-RNAs which in turn guide transgenerational gene silencing.

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Project description:Given that RHA regulates translation by binding a PCE located at the 5’ UTR of the target transcripts a genome-wide screen was performed to identify mRNAs that bind RHA in vivo. The results from duplicate experiments with samples obtained in two independent RNA immunoprecipitations identified 407 transcripts that co-immunoprecipitate with FLAG RHA. Keywords: gene expression array-based Since N-terminal FLAG tagged RHA specifically co-immunoprecipitates PCE-containing mRNAs COS cells were transfected with a CMV-FLAG-RHA construct. Cytoplasmic lysates were pooled and immunoprecipitated with anti-FLAG beads. RNA was extracted from the immunoprecipitate and used to probe rhesus macaque Affimetrix expression arrays. Steady state mRNA levels were monitored on a separate array probed with total RNA.

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Maternally inherited piRNAs silence transposons during Drosophila embryogenesis

Project description:The piRNA pathway controls transposon expression in animal germ cells, thereby ensuring genome stability over generations. piRNAs are maternally deposited and required for proper transposon silencing in adult offspring. However, a long-standing question in the field is the precise function of maternally deposited piRNAs and its associated factors during embryogenesis. Here, we probe the spatio-temporal expression patterns of several piRNA pathway components during early stages of development. Amongst those, factors required for transcriptional gene silencing (TGS) showed ubiquitous abundance in somatic and pole cells throughout the first half of embryogenesis. We further analysed the transcriptomes of various embryo stages and correlated these with the presence of selected chromatin marks. We found that a number of transposon families show bursts of transcription during early embryonic stages. Transposons heavily targeted by maternally deposited piRNAs accumulated repressive chromatin marks following their spike in expression. Furthermore, depletion of maternally deposited Piwi protein in early embryos resulted in increased expression of transposons targeted by inherited piRNAs and was accompanied by a strong loss of repressive chromatin marks at coding sequences. Overall, our data suggests a pivotal role for the piRNA pathway in transposon defence during Drosophila embryogenesis in somatic cells.

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