MRNA expression analysis in wild-type, gld-1 and cgh-1 mutant animals
Ontology highlight
ABSTRACT:
ORGANISM(S): Caenorhabditis elegans
PROVIDER: GSE33082 | GEO | 2012/06/18
SECONDARY ACCESSION(S): PRJNA155521
REPOSITORIES: GEO
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Project description:To determine mRNA expression levels in wild-type, gld-1 (q485) mutant and cgh-1(tn691) mutant animals, RNA was purified from dissected gonads of young adult worms, using the PicoPur Kit. A total of 9 samples was analyzed (triplicates of wild-type, gld-1 mutant and cgh-1 mutant animals respectively)
2012-06-17 | E-GEOD-33082 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the following subset Series: GSE33081: Identification of GLD-1 target mRNAs GSE33082: mRNA expression analysis in wild-type, gld-1 and cgh-1 mutant animals GSE33083: Polysome profiling of wild-type and gld-1 mutant animals GSE36713: mRNA expression analysis in mock and cgh-1(RNAi) animals Refer to individual Series
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Project description:To determine mRNA expression levels in mock and cgh-1(RNAi) animals, RNA was purified from young adult worms, using Trizol.
2012-06-18 | GSE36713 | GEO
Project description:To determine the translational status of mRNAs in wild-type and gld-1 mutant animals, we analyzed total and polysomal RNA levels in both strains by tiling arrays
2012-06-18 | GSE33083 | GEO
Project description:To determine the translational status of mRNAs in wild-type and gld-1 mutant animals, we analyzed total and polysomal RNA levels in both strains by tiling arrays A total of 16 samples was analyzed (quadruplicates of wild-type_total, wild-type_polysomal, gld-1_total, gld-1_polysomal RNA respectively)
2012-06-17 | E-GEOD-33083 | biostudies-arrayexpress
Project description:To determine mRNA expression levels in mock and cgh-1(RNAi) animals, RNA was purified from young adult worms, using Trizol. A total of 6 samples was analyzed (triplicates of mock and cgh-1(RNAi) animals respectively)
2012-06-17 | E-GEOD-36713 | biostudies-arrayexpress
Project description:RNA-binding proteins (RBPs) are critical regulators of gene expression and elucidating the interactions of RBPs with their RNA targets is necessary to understand how combinations of RBPs control transcriptome expression. The Quaking-related (QR) sub-family of STAR domain RBPs includes developmental regulators and tumor suppressors such as C. elegans GLD-1, which functions as a master regulator of germ line development. To understand how GLD-1 interacts with the transcriptome, we identified GLD-1 associated mRNAs by a ribonomic approach. The scale of GLD-1 mRNA interactions allowed us to determine rules governing GLD-1 target selection and to derive a predictive model where GLD-1 association with mRNA is based on the number and strength of 7-mer GLD-1 binding elements (GBEs) within UTRs. GLD-1/mRNA interactions were quantified, and predictions were verified both in vitro and in live animals, including by M-bM-^@M-^XtransplantationM-bM-^@M-^Y experiments where M-bM-^@M-^XweakM-bM-^@M-^Y and M-bM-^@M-^XstrongM-bM-^@M-^Y GBEs imposed translational repression of increasing strength on a non-target mRNA.Importantly, this study provides a unique quantitative picture of how an RBP interacts with its mRNA targets. As combinatorial regulation by multiple RBPs is thought to regulate gene expression, quantification of RBP/mRNA interactions should be a way to predict and potentially modify biological outcomes of complex posttranscriptional regulatory networks, and our analysis suggests that such an approach is possible. Wild-type (N2) or gld-1(q485); ozIs2 [gld-1::gfp::flag] worms were synchronized and harvested as young adults. To identify GLD-1 associated transcripts, two independent IP strategies were performed: 1. Comparison of anti-FLAG IP to anti-MYC IP on the gld-1(q485); ozIs2 [gld-1::gfp::flag] worm extract, 2. Comparison of anti-FLAG IP on the gld-1(q485); ozIs2 [gld-1::gfp::flag] worm extract to anti-FLAG IP on wild type extract. All IPs were performed in triplicate (12 IPs).
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Project description:To determine which mRNAs are bound by GLD-1 we performed GLD-1 and MYC (as a negative control) immunoprecipitation experiments on wildtype animals.
2012-06-18 | GSE33081 | GEO
Project description:The Poly(A)-Tail focused RNA-seq, or PAT-seq approach was utilised to determine the gene expression, poly(A)-site and polyadenylation state of the transcriptome of early C. elegans embryos having been depleated for a series of RNA binding proteins. Namely; pos-1 and mex-5 were independently knocked down using dsRNA expressing E. coli HT115(DE3) fed to ~100,000 starved/synchronized L1 larvae. When half of the population reached adulthood and half were still in the L4 stage, worms were bleached and embryos harvested and stored in Trizol. This ensured an enrichment of early embryos between 1 and 24 cell stage. mex-6(pk440) were synchronized and grown on OP50 until the same time point and embryos harvested in the same manner. Since prolonged gld-3 and gld-2 RNAi result in sterility, starved/synchronized L1 larvae were fed diluted OP50 (200uL of concentrated OP50 diluted in 2mL of M9 and starved L1s) for 16 hours. At that point the OP50 had been mostly consumed by the larvae and concentrated gld-3 or gld-2 dsRNA expressing E. coli were added to the plates. When half of the population reached adulthood and half were still in the L4 stage, worms were bleached and embryos harvested and stored in Trizol. Total RNA was isolated using standard procedures. Analysis of poly(A) dynamics in early C. elegans embryos reponding to depletion of specific RNA binding proteins and adneylation state regulators
2015-05-25 | E-GEOD-57993 | biostudies-arrayexpress
Project description:The Poly(A)-Tail focused RNA-seq, or PAT-seq approach was utilised to determine the gene expression, poly(A)-site and polyadenylation state of the transcriptome of early C. elegans embryos having been depleated for a series of RNA binding proteins. Namely; pos-1 and mex-5 were independently knocked down using dsRNA expressing E. coli HT115(DE3) fed to ~100,000 starved/synchronized L1 larvae. When half of the population reached adulthood and half were still in the L4 stage, worms were bleached and embryos harvested and stored in Trizol. This ensured an enrichment of early embryos between 1 and 24 cell stage. mex-6(pk440) were synchronized and grown on OP50 until the same time point and embryos harvested in the same manner. Since prolonged gld-3 and gld-2 RNAi result in sterility, starved/synchronized L1 larvae were fed diluted OP50 (200uL of concentrated OP50 diluted in 2mL of M9 and starved L1s) for 16 hours. At that point the OP50 had been mostly consumed by the larvae and concentrated gld-3 or gld-2 dsRNA expressing E. coli were added to the plates. When half of the population reached adulthood and half were still in the L4 stage, worms were bleached and embryos harvested and stored in Trizol. Total RNA was isolated using standard procedures.
2015-05-25 | GSE57993 | GEO