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重构异构体的挑战

在真核生物中,大多数的基因都经过可变剪接,产生多个转录异构体,大大增加了基因组的蛋白编码潜力1。来自同一个基因的可变剪接异构体可能有着明显不同、甚至拮抗的作用2。为了研究基因表达,研究人员利用新一代测序方法研究了生物体各个基因的片段,这种方法通常称为RNA测序(RNA-seq)。然而,短读长RNA-seq无法跨越全长的转录本,因而难以准确表征异构体的多样化3

无需组装,产生全长的转录本

异构体测序(Iso-Seq)可获得从转录本的5’端到poly-A尾的、全长的cDNA序列,不再需要利用异构体推断算法来重构转录组。Iso-Seq方法可提供选择性剪接外显子和转录起始位点的准确信息。对于长达10 kb的转录本,它还可提供聚腺苷酸化位点的信息,能够覆盖靶基因或整个转录组的全长异构体。

 

PacBio Sequel系统可用于高确信度地表征异构体多样性

若有意了解如何利用Iso-Seq分析转录组的复杂性,请联系我们

 References

  1. Pan, Q., et al., (2008) Deep surveying of alternative splicing complexity in the human transcriptome by high-throughput sequencing. Nature Genetics. 40, 1413-1415.
  2. Boise, LH., et al., (1993) Bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell. 74(4), 597-608.
  3. Steijger, T., et al. (2013) Assessment of transcript reconstruction methods for RNA-seq. Nature Methods. 10(12), 1177-1184.

Selected Resources