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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of wild type vs Pumilio mutant Drosophila

ABSTRACT: Total RNA was isolated with TRIZOL reagent from wild-type flies (yellow white, yw) and Pumilio mutant flies (pum13; Barker DD. et al. (1992) Genes Dev 6, 2312-26). 12 microgram of total RNA was used for cDNA synthesis using a 1:1 mixture of oligo(dT) and random nonamer primers and in the presence of amino-allyl dUTP.

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Janos Demeter

PROVIDER: E-SMDB-3532 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Wild-type versus Pumilio mutant flies

Project description:Total RNA was isolated with TRIZOL reagent from wild-type flies (yellow white, yw) and Pumilio mutant flies (pum13; Barker DD. et al. (1992) Genes Dev 6, 2312-26). 12 microgram of total RNA was used for cDNA synthesis using a 1:1 mixture of oligo(dT) and random nonamer primers and in the presence of amino-allyl dUTP. Set of arrays that are part of repeated experiments Keywords: Biological Replicate

2006-02-28 | GSE3580 | GEO

Wild-type versus Pumilio mutant flies

2010-07-01 | E-GEOD-3580 | biostudies-arrayexpress

Transcription profiling of adult fly vs embryo (0-16h), Gerber AP. 2005

Project description:Total RNA was isolated from wild-type adult flies (yellow white) and from embryos (cs, 0-16h) with TRIZOL reagent. 12 microgram of each total RNA were labeled with fluorescent dyes, following cDNA synthesis with amino-allyl dUTP in addition to the four natural dNTPs using a 1:1 mixture of oligo(dT) and random nonamer primers.

2006-11-11 | E-SMDB-3536 | biostudies-arrayexpress

Adult fly versus embryo (0-16h)

2006-02-28 | GSE3581 | GEO

Adult fly versus embryo (0-16h)

2010-07-01 | E-GEOD-3581 | biostudies-arrayexpress

Oophaga pumilio

Project description:Oophaga pumilio overview

| PRJNA550473 | ENA

Rhabdomys pumilio

Project description:Rhabdomys pumilio Genome sequencing

| PRJNA399358 | ENA

Oophaga pumilio

Project description:Oophaga pumilio whole genome sequencing

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Gene expression profiles in NORAD knockout and PUMILIO overexpressing cells

Project description:Purpose: PUMILIO proteins are known to repress target genes by specifically binding to PUMILIO response elements (PREs) in target mRNAs. NORAD is a noncoding RNA that negatively regulates PUMILIO activity. The goal of this study was to determine the gene expression changes that result from knockout of NORAD or overexpression of PUMILIO and to test whether NORAD knockout causes PUMILIO hyperactivity. Methods: RNA-seq libraries were prepared using the TruSeq Stranded Total RNA with Ribo-Zero Human/Mouse/Rat Sample Preparation kit (Illumina) and sequenced using the 100 bp paired-end protocol on an Illumina HiSeq 2000. For comparing NORAD+/+ and NORAD-/- HCT116 cells, 3 biological replicates per genotype were sequenced. For PUM overexpression experiments, 3 replicates of GFP-expressing HCT116 cells (negative control) and 2 independent PUM1- or PUM2-overexpressing clones (2 replicates each) were sequenced. Results: Gene expression profiles show that PUMILIO target genes are downregulated in both NORAD knockout cells and PUMILIO overexpressing cells. Conclusions: These data indicate that NORAD sequesters PUMILIO, preventing excessive repression of PUMILIO target genes that are important for maintaining genomic stability.

2015-12-03 | GSE75440 | GEO

Regulation of the Drosophila transcriptome by Pumilio and CCR4-NOT deadenylase

Project description:The sequence-specific RNA-binding protein Pumilio controls development of Drosophila; however, the network of mRNAs that it regulates remains incompletely characterized. In this study, we utilize knockdown and knockout approaches coupled with RNA-Seq to measure the impact of Pumilio on the transcriptome of Drosophila cells. We also used an improved RNA co-immunoprecipitation method to identify Pumilio bound mRNAs in Drosophila embryos. Integration of these datasets with the content of Pumilio binding motifs across the transcriptome revealed novel direct Pumilio target genes involved in neural, muscle, wing, and germ cell development, and cellular proliferation. These genes include components of Wnt, TGF-beta, MAPK/ERK, and Notch signaling pathways, DNA replication, and lipid metabolism. Additionally, we identified the mRNAs regulated by the CCR4-NOT deadenylase complex, a key factor in Pumilio-mediated repression, and observed concordant regulation of Pumilio:CCR4-NOT target mRNAs. Computational modeling revealed that Pumilio binding site location, number, density, and context are important determinants of regulation. Moreover, the content of optimal synonymous codons exhibits a striking functional relationship to Pumilio and CCR4-NOT regulation, indicating that the inherent translation efficiency and stability of the mRNA modulates their response to these trans-acting regulatory factors. Together, the results of this work provide new insights into the Pumilio regulatory network and mechanisms, and the parameters that influence the efficacy of Pumilio-mediated regulation.

2023-09-03 | GSE241940 | GEO

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