Global Splicing Pattern Reversion during Somatic Cell Reprogramming
Global Splicing Pattern Reversion during Somatic Cell Reprogramming
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DOI：
10.1016/j.celrep.2013.09.016
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发表时间：
2013-10-01
期刊：
影响因子：
8.8
通讯作者：
Yamamoto, Takuya
中科院分区：
文献类型：
--
作者：
Ohta, Sho;Nishida, Eisuke;Yamamoto, Takuya
选择性剪接从单个基因产生多个转录本，细胞类型特异性剪接特征对于细胞的特性和功能很重要。最近，体细胞已被证明在转录因子的强制表达后会发生去分化。然而，目前尚不清楚体细胞剪接是否在重编程过程中进行重组。在这里，通过将深度测序与高通量绝对 qRT-PCR 相结合，我们表明体细胞剪接图谱在重编程过程中恢复为多能剪接图谱。值得注意的是，多能干细胞中的剪接模式与睾丸中的剪接模式相似，并且调节区在长度和序列上具有特定的特征。此外，我们的 siRNA 筛选还鉴定出了调节 iPSC 剪接事件的 RNA 结合蛋白。然后我们证明了两种 RNA 结合蛋白 U2af1 和 Srsf3 在体细胞重编程中发挥作用。我们的结果表明，剪接的剧烈改变代表了参与重编程过程的分子网络的一部分。
Alternative splicing generates multiple transcripts from a single gene, and cell-type-specific splicing profiles are important for the properties and functions of the cells. Recently, somatic cells have been shown to undergo dedifferentiation after the forced expression of transcription factors. However, it remains unclear whether somatic cell splicing is reorganized during reprogramming. Here, by combining deep sequencing with high-throughput absolute qRT-PCR, we show that somatic splicing profiles revert to pluripotent ones during reprogramming. Remarkably, the splicing pattern in pluripotent stem cells resembles that in testes, and the regulatory regions have specific characteristics in length and sequence. Furthermore, our siRNA screen has identified RNA-binding proteins that regulate splicing events in iPSCs. We have then demonstrated that two of the RNA-binding proteins, U2af1 and Srsf3, play a role in somatic cell reprogramming. Our results indicate that the drastic alteration in splicing represents part of the molecular network involved in the reprogramming process.