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Developmental timecourse of C. elegans small RNAs with 5p monophosphates

ABSTRACT: These datasets profile the expression of small RNAs with 5p monophosphates and 3p hydroxyls (including miRNAs, 21U-RNAs) across C. elegans development and in dauer, glp-4 mutant, and mixed-stage wt worms. These datasets were prepared in the David P. Bartel laboratory using T4 RNA ligase Rnl2(1-249). Keywords: miRNA and 21U-RNA discovery and developmental expression profiling CE-devProfile-5pDependent-Illumina 1 flowcell each for Embryo, L1, L2, L3, L4, Adult, Dauer, Glp-4 mutant; 6 flowcells mixed stage

ORGANISM(S): Caenorhabditis elegans

SUBMITTER: James Ruby

PROVIDER: E-GEOD-11734 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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PRG-1 and 21U-RNAs interact to form the piRNA complex required for fertility in C. elegans.

Batista Pedro J PJ Ruby J Graham JG Claycomb Julie M JM Chiang Rosaria R Fahlgren Noah N Kasschau Kristin D KD Chaves Daniel A DA Gu Weifeng W Vasale Jessica J JJ Duan Shenghua S Conte Darryl D Luo Shujun S Schroth Gary P GP Carrington James C JC Bartel David P DP Mello Craig C CC

Molecular cell 20080619 1

In metazoans, Piwi-related Argonaute proteins have been linked to germline maintenance, and to a class of germline-enriched small RNAs termed piRNAs. Here we show that an abundant class of 21 nucleotide small RNAs (21U-RNAs) are expressed in the C. elegans germline, interact with the C. elegans Piwi family member PRG-1, and depend on PRG-1 activity for their accumulation. The PRG-1 protein is expressed throughout development and localizes to nuage-like structures called P granules. Although 21U- ...[more]

PMID: 18571452

Similar Datasets

Mutant analysis of C. elegans small RNAs with 5p monophosphates

Project description:These datasets profile the expression of small RNAs with 5p monophosphates and 3p hydroxyls (including miRNAs, 21U-RNAs) in young adult worms from the wt, prg-1 mutant, and fog-2 mutant backgrounds. These datasets were prepared in the Craig C. Mello laboratory using T4 RNA ligase 1. Keywords: miRNA and 21U-RNA discovery and mutant expression profiling CE-mutProfile-5pDependent-Illumina 1 flowcell each for the wt, prg-1 mutant, and fog-2 mutant

2008-06-15 | E-GEOD-11735 | biostudies-arrayexpress

Small-RNAs in L4 and young adult stages

Project description:Small endogenous C. elegans RNAs from L4 and young adult worms were prepared for sequencing using a protocol derived from Batista et al., (2008) and Lau et al. (2001). The small-RNA libraries were constructed using a method that does not require a 5M-bM-^@M-^Y monophosphate (called 5M-bM-^@M-^Y monophosphate-independent method, Ambros et al., 2003) to profile secondary siRNAs that have 5M-bM-^@M-^Y triphosphorylated G. All preprocessed small-RNA reads were mapped to genome (ce6), allowing no mismatches. After excluding miRNAs, 21U RNAs, rRNAs, and other structural ncRNAs, the remaining reads were classified as 22G RNAs, 26G RNAs, and other siRNAs, based on their length and 5M-bM-^@M-2 terminal nucleotide. Small-RNA libraries were sequenced in L4 and young adult stages in C.elegans.

2012-06-12 | E-GEOD-36394 | biostudies-arrayexpress

C. elegans piRNAs mediate the genome-wide surveillance of germline transcripts

Project description:Piwi Argonautes and Piwi-interacting RNAs (piRNAs) mediate genome defense by targeting transposons. However, many piRNA species lack obvious sequence complementarity to transposons or other loci. For example, only one C. elegans transposon is a known piRNA target. Here we show that, in mutants lacking the Piwi Argonaute PRG-1 and associated piRNAs (21U-RNAs), many silent loci in the germline exhibit increased levels of mRNA expression and depletion of an amplified RNAdependent RNA polymerase (RdRP)-derived species of small RNA termed 22G-RNAs. Sequences depleted of 22G-RNAs are enriched nearby potential target sites that base pair imperfectly but extensively to 21U-RNAs. We show that PRG-1 is required to initiate, but not to maintain, silencing of transgenes engineered to contain complementarity to endogenous 21U-RNAs. Our findings support a model in which C. elegans piRNAs utilize their enormous repertoire of targeting capacity to scan the germline transcriptome for foreign sequences, while endogenous germline-expressed genes are actively protected from piRNA-induced silencing. Examine small RNA population changes in prg-1 and rescued strains

2012-09-01 | E-GEOD-38723 | biostudies-arrayexpress

Developmental timecourse of C. elegans small RNAs with 5p monophosphates

Project description:These datasets profile the expression of small RNAs with 5p monophosphates and 3p hydroxyls (including miRNAs, 21U-RNAs) across C. elegans development and in dauer, glp-4 mutant, and mixed-stage wt worms. These datasets were prepared in the David P. Bartel laboratory using T4 RNA ligase Rnl2(1-249). Keywords: miRNA and 21U-RNA discovery and developmental expression profiling

2008-06-16 | GSE11734 | GEO

A Genome-wide RNAi screen identifies factors required for distinct stages of C. elegans piRNA biogenesis

Project description:In animals, piRNAs, and their associated Piwi proteins, guard germ cell genomes against mobile genetic elements via an RNAi-like mechanism. In C. elegans, 21U-RNAs comprise the piRNA class and these collaborate with 22G RNAs, via unclear mechanisms, to discriminate self from non-self and selectively and heritably silence the latter. Recent work indicates that 21U-RNAs are post-transcriptional processing products of individual transcription units that produce ~26 nucleotide capped precursors. Yet, nothing is known of how the expression of precursors is controlled or of how primary transcripts give rise to mature small RNAs. We conducted a genome-wide RNAi screen to identify components of the 21U biogenesis machinery. Screening by direct, qPCR-based measurements of mature 21U-RNA levels, we identified 22 genes important for 21U-RNA production, termed TOFUs (Twenty-One-u Fouled Up). We also identified 7 genes that normally repress 21U production. By measuring mature 21U-RNA and precursor levels for the 7 strongest hits from the screen, we have assigned factors to discrete stages of 21U-RNA production. Our work has identified factors separately required for the transcription of 21U precursors, and the processing of these precursors into mature 21U-RNAs, thereby providing an essential resource for studying the biogenesis of this important small RNA class. Small RNA and capped small RNA sequencing from total RNA of C. elegans subjected to different RNAi and different C. elegans mutants

2014-04-01 | E-GEOD-53970 | biostudies-arrayexpress

Mutant analysis of C. elegans small RNAs, 5p-end-independent

Project description:These datasets profile the expression of small RNAs with 3p hydroxyls (including endo-siRNAs, miRNAs, and 21U-RNAs) in young adult worms from the wt and prg-1 mutant backgrounds. They were prepared using an approach that captures small RNAs independent of the covalent status of their 5p termini, as described in Ambros et al. (2003) Curr Biol 13:807-18. These datasets were prepared in the Craig C. Mello laboratory. Keywords: endo-siRNA mutant expression profiling CE-prg1-endoSiRNAs-Illumina 1 flowcell each for the wt and prg-1 mutant

2008-06-15 | E-GEOD-11736 | biostudies-arrayexpress

Argonautes promote male fertility and provide a paternal memory of germline gene expression in C. elegans

Project description:During each life cycle germ cells preserve and pass on both genetic and epigenetic information. In C. elegans, the ALG-3/4 Argonaute (AGO) proteins and their small-RNA cofactors are expressed during male gametogenesis and promote male fertility. Here we show that the CSR-1 AGO functions with ALG-3/4 to positively regulate target genes required for spermiogenesis. Our findings suggest that ALG-3/4 functions during spermatogenesis to amplify a small-RNA signal that represents an epigenetic memory of male-specific gene expression, while CSR-1, which is abundant in mature sperm, transmits this memory to offspring. Surprisingly, in addition to small RNAs targeting male-specific genes, we show that males also harbor an extensive repertoire of CSR-1 small RNAs targeting oogenesis-specific mRNAs. Together these findings suggest that C. elegans sperm transmit not only the genome but also epigenetic binary signals in the form of Argonaute/small-RNA complexes that constitute a memory of which genes were active in preceding generations. Examine small RNA changes in WT and alg-3/4 mutant males cultured at 20M-BM-0C and 25M-BM-0C, as well as determine the small RNAs enriched in a FLAG::CSR-1 IP from male worms grown at 25M-BM-0C. mRNA sequencing was also performed to determine how transcripts targeted by small RNAs change in mutant background at 20M-BM-0C and 25M-BM-0C.

2013-12-19 | E-GEOD-49672 | biostudies-arrayexpress

Connecting microRNA genes to the core transcriptional regulatory circuitry of embryonic stem cells

Project description:MicroRNAs (miRNAs) are crucial for normal embryonic stem (ES) cell self-renewal and cellular differentiation, but how miRNA gene expression is controlled by the key transcriptional regulators of ES cells has not been established. We describe here a new map of the transcriptional regulatory circuitry of ES cells that incorporates both protein-coding and miRNA genes, and which is based on high-resolution ChIP-seq data, systematic identification of miRNA promoters, and quantitative sequencing of short transcripts in multiple cell types. We find that the key ES cell transcription factors are associated with promoters for most miRNAs that are preferentially expressed in ES cells and with promoters for a set of silent miRNA genes. This silent set of miRNA genes is co-occupied by Polycomb Group proteins in ES cells and expressed in a tissue-specific fashion in differentiated cells. These data reveal how key ES cell transcription factors promote the miRNA expression program that contributes to self-renewal and cellular differentiation, and integrate miRNAs and their targets into an expanded model of the regulatory circuitry controlling ES cell identity. Keywords: ChIP-seq analysis of ES cell transcriptional regulators and chromatin modifications. Cell-type comparison of short RNA transcritome. Analysis of changes in short RNA transcritome upon Oct4 ablation. ChIP-seq in murine embryonic stem cells for Oct4, Sox2 (2 runs), Nanog (2 runs), Tcf3 (2 runs), Suz12 (2 runs), H3K4me3 (4 runs), H3k79me2 (2 runs), H3k36me3 (2 runs) and whole cell extract input DNA (WCE, 2 runs). Short transcript sequencing from murine embryonic stem cells (mES, v6.5), mouse embryonic fibroblasts (MEF), murine neural precursor cells (NPC), and ZHBT-c4 cells (from Austin Smith) untreated (0h), with 12 hours of doxycyclin treatment (12h), and with 24 hours of doxycyclin treatment (24h).

2008-07-29 | E-GEOD-11724 | biostudies-arrayexpress

The Translin-TRAX complex/C3PO is a ribonuclease in tRNA processing

Project description:The endogenous RNA substrates of Translin-TRAX complexes (also known as C3POs) and how they regulate diverse biological processes remain unknown. Here we show that Translin and TRAX do not play a significant role in RNAi in the filamentous fungus Neurospora crassa. Instead, the Neurospora C3PO complex functions as a ribonuclease that removes the 5M-bM-^@M-^Y pre-tRNA fragments after the processing of pre-tRNAs by RNase P. In translin and trax mutants, 5M-bM-^@M-^Y pre-tRNA fragments accumulate to very high levels that can be degraded specifically by both recombinant and endogenous Neurospora C3PO and recombinant Drosophila C3PO. In addition, the mutants have elevated tRNA levels and increased levels of protein translation and are more resistant to a programmed cell-death inducing agent. Together, this study identified the endogenous RNA substrates of C3PO and provides a potential explanation for its roles in seemingly diverse biological processes. Examine small RNA population changes in two different strain background

2012-07-30 | E-GEOD-38334 | biostudies-arrayexpress

High-throughput pyrosequencing of endogenous small RNAs from CSR-1 IP complexes and csr-1(tm892) and ego-1(om97) mutants

Project description:High-throughput pyrosequencing of endogenous small RNAs from CSR-1 IP complexes and csr-1(tm892) and ego-1(om97) mutants with corresponding controls. RNAi-related pathways regulate diverse processes, from developmental timing to transposon silencing. Here, we show that in C. elegans the Argonaute CSR-1, the RNA-dependent RNA polymerase EGO-1, the Dicer-related helicase DRH-3, and the Tudor-domain protein EKL-1 localize to chromosomes and are required for proper chromosome segregation. In the absence of these factors chromosomes fail to align at the metaphase plate and kinetochores do not orient to opposing spindle poles. Surprisingly, the CSR-1 interacting small RNAs (22G-RNAs) are antisense to thousands of germline-expressed protein-coding genes. Nematodes assemble holocentric chromosomes in which continuous kinetochores must span the expressed domains of the genome. We show that CSR-1 interacts with chromatin at target loci, but does not down-regulate target mRNA or protein levels. Instead, our findings support a model in which CSR-1 complexes target protein-coding domains to promote their proper organization within the holocentric chromosomes of C. elegans. 5 samples examined. Small RNAs that co-immunopercipitate with CSR-1 protein and input sample. Small RNAs from csr-1(tm892) and ego-1(om97) mutants and corresponding congenic wild type strain.

2009-10-02 | E-GEOD-18165 | biostudies-arrayexpress

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