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SRX815493: GSM1567913: Adrenal_18wk; Homo sapiens; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 29.6M spots, 5.9G bases, 4Gb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Tissue-specific circular RNA induction during human fetal development
show Abstracthide Abstract
The pervasive expression of circular RNA from protein coding loci is a recently discovered feature of many eukaryotic gene expression programs. Computational methods to discover and quantify circular RNA are essential to the study of the mechanisms of circular RNA biogenesis and potential functional roles they may play. In this paper, we present a new statistical algorithm that increases the sensitivity and specificity of circular RNA detection.by discovering and quantifying circular and linear RNA splicing events at both annotated exon boundaries and in un-annotated regions of the genome Unlike previous approaches which rely on heuristics like read count and homology between exons predicted to be circularized to determine confidence in prediction of circular RNA expression, our algorithm is a statistical approach. We have used this algorithm to discover general induction of circular RNAs in many tissues during human fetal development. We find that some regions of the brain show marked enrichment for genes where circular RNA is the dominant isoform. Beyond this global trend, specific circular RNAs are tissue specifically induced during fetal development, including a circular isoform of NCX1 in the developing fetal heart that, by 20 weeks, is more highly expressed than the linear isoform as well as beta-actin. In addition, while the vast majority of circular RNA production occurs at canonical U2 (major spliceosome) splice sites, we find the first examples of developmentally induced circular RNAs processed by the U12 (minor) spliceosome, and an enriched propensity of U12 donors to splice into circular RNA at un-annotated, rather than annotated, exons. Together, our algorithm and its results suggest a potentially significant role for circular RNA in human development. Overall design: 35 human fetal samples from 6 tissues (3 - 7 replicates per tissue) collected between 10 and 20 weeks gestational time were sequenced using Illumina TruSeq Stranded Total RNA with Ribo-Zero Gold sample prep kit.
Sample: GEO accession GSM1567913 is currently private and is scheduled to be released on Dec 31, 2015.
SAMN03267761 • SRS795839 • All experiments • All runs
Organism: Homo sapiens
Library:
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Fetal tissue samples <100 mg in size were placed into DNase- and RNase-free 1.5 ml microfuge tubes containing 1 ml of RNAlater RNA Stabilization Reagent (Qiagen) within 1 hour of the pregnancy termination procedure. After storage at room temperature for a period of 24-72 h, excess RNAlater was removed from the microfuge tubes and the samples were placed in the -80oC freezer for storage until RNA isolation was performed. The tissues were lysed in mirVana (Life Technologies) lysis buffer, using a Mini-Beadbeater-16 (Biospec), with agitation for 1 min in the presence of 1 mm zirconia beads. Samples were then centrifuged at maximum speed in a tabletop microcentrifuge for 1 min and the lysed solution was transferred to a fresh microfuge tube. The remainder of the extraction was per the manufacturer's protocol for the mirVana kit (Life Technologies). Following RNA isolation (mirVana miRNA Isolation Kit, Life Technologies, Inc.), the RNA was quantified (Qubit RNA Assay Kit, Life Technologies, Inc.), and quality controlled (RNA6000 Nano Kit and BioAnalyzer 2100, Agilent). 200 ng to 1000 ng was used as input for the Illumina TruSeq Stranded Total RNA with Ribo-Zero Gold sample prep kit (Illumina, Inc.) and sequencing libraries were created according to the manufacturer’s protocol. Briefly, first both cytoplasmic and mitochondrial rRNA was removed by selectively hybridizing biotinylated probes to target sequences and using magnetic beads to capture the bound products. Following rRNA removal, the RNA was fragmented and copied into first strand cDNA using random primers and reverse transcriptase. Next, second strand cDNA synthesis was completed using DNA Polymerase I and RNase H. The cDNA was then ligated to Illumina supplied adapters and enriched with PCR to create the final cDNA libraries. The libraries were then pooled and sequenced on a HiSeq 2000 (Illumina, Inc.) instrument as per manufacturer’s instructions. Sequencing was performed up to 2 X 101 cycles.
Experiment attributes:
GEO Accession: GSM1567913
Links:
External link:
Runs: 1 run, 29.6M spots, 5.9G bases, 4Gb
Run# of Spots# of BasesSizePublished
SRR172128129,614,2805.9G4Gb2015-06-25

ID:
1165194

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