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SRX348499: GSM1229062: g_SF_iPS_3_rep1; Gorilla gorilla; RNA-Seq
1 ILLUMINA (Illumina Genome Analyzer IIx) run: 4.2M spots, 321.2M bases, 241.7Mb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Primate iPS cells as tools for evolutionary analyses
show Abstracthide Abstract
Induced pluripotent stem cells (iPSCs) are regarded as a central tool to understand human biology in health and disease. Similarly, iPSCs from closely related species should be a central tool to understand human evolution and to identify conserved and variable patterns of iPSC disease models. Here, we have generated human, gorilla, bonobo and cynomolgus monkey iPSCs. We show that these cells are well comparable in their differentiation potential and generally similar to human, cynomolgus and rhesus monkey embryonic stem cells (ESCs). RNA sequencing reveals that expression differences among clones, individuals and stem cell type are all of very similar magnitude within a species. In contrast, expression differences between closely related primate species are three times larger and most genes show significant expression differences among the analysed species. However, pseudogenes differ more than twice as much, suggesting that evolution of expression levels in primate stem cells is rapid, but constrained. These patterns in pluripotent stem cells are comparable to those found in other tissues except testis. Hence, primate iPSCs reveal insights into general primate gene expression evolution and should provide a rich source to identify conserved and species-specific gene expression patterns for cellular phenotypes. Contributors: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, D-04103 Leipzig, Germany Overall design: We used expression profiling to characterize five gorilla, two bonobo and three macaque iPS clones as well as three iPS clones from two human individuals, three human embryonic stem (ES) cell lines and three macaque ES cell lines. We generated tagged RNA-Seq libraries from these 19 samples including four technical replicates (23 samples). Over 100 million single end reads were generated on the Illumina platform.
Sample: g_SF_iPS_3_rep1
SAMN02353893 • SRS479275 • All experiments • All runs
Organism: Gorilla gorilla
Library:
Instrument: Illumina Genome Analyzer IIx
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Cells were grown as described above in 6-well plates, removed from mouse feeder cells as good as possible and cells from one plate suspended in 1ml Trizol (Invitrogen). Total RNA from 500µl Trizol was isolated and further purified using RNAeasy columns (Quiagen). RNA-Seq libraries were constructed using a custom protocol combining the Illumina TruSeq RNA protocol to generate cDNA and a double-indexing protocol [PMID: 22021376, PMID: 20516186] to generate sequencing libraries from this cDNA (full detailed protocol available upon request). Briefly, 1µg of total RNA was mRNA purified twice (Sera-Mag Magnetic Oligo(dT) beads, Thermo Scientific), cleaned up using RNAClean beads (Beckman Coulter), eluted in 2µl first strand buffer and 0.5µl random primers (Invitrogen) and fragmented for 7 min at 85°C. After first and second strand synthesis, cDNA was end-repaired, custom P5 and P7 adapters were ligated, blunted, quantified and amplified in an indexing PCR essentially as described [PMID: 20516186], except that both ends were indexed [PMID: 22021376]. The quality and concentration of the library were assessed on a BioAnalyzer.
Experiment attributes:
GEO Accession: GSM1229062
Links:
External link:
Runs: 1 run, 4.2M spots, 321.2M bases, 241.7Mb
Run# of Spots# of BasesSizePublished
SRR9761794,226,045321.2M241.7Mb2014-02-19

ID:
494909

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