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SRX2788642: GSM2604565: pol2chipseq.cuffmut.rep1; Drosophila melanogaster; ChIP-Seq
1 ILLUMINA (Illumina Genome Analyzer II) run: 127.2M spots, 11.5G bases, 7.7Gb downloads

Submitted by: NCBI (GEO)
Study: Integrative analysis unveils new functions for the Drosophila Cutoff protein in non-coding RNA biogenesis and gene regulation
show Abstracthide Abstract
Piwi-Interacting RNAs (piRNAs) are central components of the piRNA pathway, which directs transposon silencing and guarantees genome integrity in the germ cells of several metazoans. In Drosophila, piRNAs are produced from discrete regions of the genome termed piRNA clusters, whose expression relies on the RDC complex comprised of the core proteins Rhino, Deadlock and Cutoff. To date, the RDC complex has been exclusively implicated in the regulation of the piRNA loci. Here we further elucidate the function of Cutoff and the RDC complex by performing genome-wide ChIP-seq and RNA-seq assays in the Drosophila germline and analyzing these data together with other publicly available data sets. In agreement with previous studies, we confirm that Cutoff is involved in the transcriptional regulation of piRNA clusters and in the repression of transposable elements in germ cells. Surprisingly, however, we find that Cutoff is enriched at and affects the expression of other non-coding RNAs, including spliceosomal RNAs (snRNAs) and small nucleolar RNAs (snoRNAs). At least in some instances, Cutoff appears to act at a transcriptional level in concert with Rhino and perhaps Deadlock. Finally, we show that mutations in Cutoff result in the deregulation of hundreds of protein-coding genes in germ cells. Our study uncovers a broader function for the RDC complex in the Drosophila germline development. Overall design: Cutoff ChIP-seq (2 replicates with ChIP, 3 replicates with control), Pol2 ChIP-seq (3 replicates in WT, 3 replicates in cuff mutant), RNA-seq (3 replicates in WT, 3 replicates in cuff mutant)
Sample: pol2chipseq.cuffmut.rep1
SAMN06909689 • SRS2170205 • All experiments • All runs
Library:
Instrument: Illumina Genome Analyzer II
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: RNA-seq and ChIP-seq assays were performed in triplicate and for each biological replicate we dissected ovaries from approximately 30 female flies (2-4 days old). Total RNA was extracted using Trizol reagents (Life Technologies) and treated with turbo DNase-free kit (Ambion) to eliminate traces. of genomic DNA. DNA-free total RNA was subjected to two rounds of rRNA depletion using the Ribo-Zero rRNA removal kit (Epicentre) as per the manufacturer's instructions. The same ChIP protocol was employed for the ChIP-seq assays as previously described (Pane et al. 2011; Blythe et al. 2009). The Cuff-EGFP expressing transgenic line, GFP antibodies and the conditions used for the Cuff ChIP-seq experiment were previously described (Pane et al. 2011; Blythe et al. 2009). The RNA-seq library preparation was obtained with the ScriptSeq v2 kit (Epicentre) and the libraries were sequenced on the Illumina platforms. Chromatin fragments were used to generate DNA libraries with the ChIP-seq DNA Sample Prep kit (Illumina).
Experiment attributes:
GEO Accession: GSM2604565
Links:
Runs: 1 run, 127.2M spots, 11.5G bases, 7.7Gb
Run# of Spots# of BasesSizePublished
SRR5515262127,170,27411.5G7.7Gb2017-05-10

ID:
4022560

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