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Ultra-deep profiling of alternatively spliced Drosophila Dscam isoforms by circularization-assisted multi-segment sequencing

Official URL:https://doi.org/10.1038/emboj.2013.144
PubMed:View item in PubMed
Creators Name:Sun, W. and You, X. and Gogol-Doering, A. and He, H. and Kise, Y. and Sohn, M. and Chen, T. and Klebes, A. and Schmucker, D. and Chen, W.
Journal Title:EMBO Journal
Journal Abbreviation:EMBO J
Page Range:2029-2038
Date:17 July 2013
Keywords:Alternative Splicing, CAMSeq, Dscam, Ultra-Deep Profiling, Animals, Drosophila Melanogaster
Abstract:The Drosophila melanogaster gene Dscam (Down syndrome cell adhesion molecule) can generate thousands of different ectodomains via mutual exclusive splicing of three large exon clusters. The isoform diversity plays a profound role in both neuronal wiring and pathogen recognition. However, the isoform expression pattern at the global level remained unexplored. Here, we developed a novel method that allows for direct quantification of the alternatively spliced exon combinations from over hundreds of millions of Dscam transcripts in one sequencing run. With unprecedented sequencing depth, we detected a total of 18 496 isoforms, out of 19 008 theoretically possible combinations. Importantly, we demonstrated that alternative splicing between different clusters is independent. Moreover, the isoforms were expressed across a broad dynamic range, with significant bias in cell/tissue and developmental stage-specific patterns. Hitherto underappreciated, such bias can dramatically reduce the ability of neurons to display unique surface receptor codes. Therefore, the seemingly excessive diversity encoded in the Dscam locus might nevertheless be essential for a robust self and non-self discrimination in neurons.
Publisher:Nature Publishing Group (U.S.A.)
Item Type:Article

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