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首页> 外文期刊>EMBO Journal >Ultra-deep profiling of alternatively spliced Drosophila Dscam isoforms by circularization-assisted multi-segment sequencing
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Ultra-deep profiling of alternatively spliced Drosophila Dscam isoforms by circularization-assisted multi-segment sequencing

机译：超深或者拼接的剖析果蝇Dscam亚型,circularization-assisted为多段测序

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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.
机译：黑腹果蝇基因Dscam(下降症细胞粘附分子)可以生成成千上万的不同ectodomains通过相互的独家的三大集群外显子剪接。同种型多样性起着深远的作用神经连接和病原体识别。然而,在同种型表达模式全球层面上仍然是未知的。开发了一种新方法,允许直接量化的或者外显子拼接的从成千上万的组合Dscam记录在一个序列运行。前所未有的测序深度,我们检测到一个496年共有18亚型,19 008理论上可能的组合。我们表明,可变剪接不同的集群是独立的。亚型表达在一个广泛的动态范围,细胞/组织和明显的偏见发展stage-specific模式。人的重视,这样的偏见会显著减少神经元显示独特的能力表面受体编码。过度多元化Dscam轨迹编码不过可能是必不可少的一个健壮的自我在神经元和异己分子歧视。

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