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mRNA-seq with agnostic splice site discovery for nervous system transcriptomics tested in chronic pain

Title:
mRNA-seq with agnostic splice site discovery for nervous system transcriptomics tested in chronic pain
Abstract:
mRNA-seq is a paradigm-shifting technology because of its superior sensitivity and dynamic range and its potential to capture transcriptomes in an agnostic fashion, i.e., independently of existing genome annotations. Implementation of the agnostic approach, however, has not yet been fully achieved. In particular, agnostic mapping of pre-mRNA splice sites has not been demonstrated. The present study pursued dual goals: (1) to advance mRNA-seq bioinformatics toward unbiased transcriptome capture and (2) to demonstrate its potential for discovery in neuroscience by applying the approach to an in vivo model of neurological disease. We have performed mRNA-seq on the L4 dorsal root ganglion (DRG) of rats with chronic neuropathic pain induced by spinal nerve ligation (SNL) of the neighboring (L5) spinal nerve. We found that 12.4% of known genes were induced and 7% were suppressed in the dysfunctional (but anatomically intact) L4 DRG 2 wk after SNL. These alterations persisted chronically (2 mo). Using a read cluster classifier with strong test characteristics (ROC area 97%), we discovered 10,464 novel exons. A new algorithm for agnostic mapping of pre-mRNA splice junctions (SJs) achieved a precision of 97%. Integration of information from all mRNA-seq read classes including SJs led to genome reannotations specifically relevant for the species used (rat), the anatomical site studied (DRG), and the neurological disease considered (pain); for example, a 64-exon coreceptor for the nociceptive transmitter substance P was identified, and 21.9% of newly discovered exons were shown to be dysregulated. Thus, mRNA-seq with agnostic analysis methods appears to provide a highly productive approach for in vivo transcriptomics in the nervous system.
Year:
2010
Publication type:
Journal article
Journal:
Genome Research
ISSN:
1549-5469
Number:
6
Volume:
20
Pages:
847-860
Language:
English
Document status:
Open access
PubListerURL:
https://publister.bib.th-wildau.de/publister/public/publication/553
Hammer, P., Banck, M., Amberg, R., Wang, C., Petznick, G., Luo, S., et al. (2010). mRNA-seq with agnostic splice site discovery for nervous system transcriptomics tested in chronic pain Genome Research. 20 (6), 847-860.
@article{553,
    author           = {Hammer, Paul and Banck, Michaela S. and Amberg, Ronny and Wang, Cheng and Petznick, Gabriele and Luo, Shujun and Khrebtukova, Irina and Schroth, Gary P. and Beyerlein, Peter and Beutler, Andreas S.},
    title            = {mRNA-seq with agnostic splice site discovery for nervous system transcriptomics tested in chronic pain},
    journal          = {Genome Research},
    year             = {2010},
    volume           = {20},
    number           = {6},
    pages            = {847-860},
    doi              = {10.1101/gr.101204.109},
}


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