Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs.
Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs.
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DOI：
10.1038/s41467-018-02866-0
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发表时间：
2018-02-12
影响因子：
16.6
通讯作者：
Nikaido I
中科院分区：
文献类型：
--
作者：
Hayashi T;Ozaki H;Sasagawa Y;Umeda M;Danno H;Nikaido I
总RNA测序已用于揭示多聚（A）和非多聚（A）RNA表达、RNA加工和增强子活性。迄今为止，尽管单细胞生物学技术具有潜力，但还没有开发出单细胞全长总RNA测序的方法。在这里，我们描述了随机置换扩增测序（RamDA-seq），这是第一种用于单细胞的全长总RNA测序方法。与其他方法相比，RamDA-seq显示出对非poly（A）RNA的高灵敏度和几乎完全的全长转录物覆盖。使用RamDA-seq与小鼠胚胎干细胞的分化时程样本，我们揭示了数百种动态调节的非poly（A）转录本，包括组蛋白转录本和长链非编码RNA Neat 1。此外，RamDA-seq分析>300-kb内含子中的递归剪接。RamDA-seq还检测增强子RNA及其在单细胞中的细胞类型特异性活性。总之，我们证明RamDA-seq可以帮助研究单细胞中基因表达，RNA加工事件和转录调控的动态。总RNA测序已用于分析多聚（A）和非多聚（A）RNA表达、加工和增强子的活性。在这里，作者开发了RamDA-seq，一种在单细胞中进行全长总RNA测序的方法。
Total RNA sequencing has been used to reveal poly(A) and non-poly(A) RNA expression, RNA processing and enhancer activity. To date, no method for full-length total RNA sequencing of single cells has been developed despite the potential of this technology for single-cell biology. Here we describe random displacement amplification sequencing (RamDA-seq), the first full-length total RNA-sequencing method for single cells. Compared with other methods, RamDA-seq shows high sensitivity to non-poly(A) RNA and near-complete full-length transcript coverage. Using RamDA-seq with differentiation time course samples of mouse embryonic stem cells, we reveal hundreds of dynamically regulated non-poly(A) transcripts, including histone transcripts and long noncoding RNA Neat1. Moreover, RamDA-seq profiles recursive splicing in >300-kb introns. RamDA-seq also detects enhancer RNAs and their cell type-specific activity in single cells. Taken together, we demonstrate that RamDA-seq could help investigate the dynamics of gene expression, RNA-processing events and transcriptional regulation in single cells. Total RNA sequencing has been used to profile poly(A) and non-poly(A) RNA expression, processing and the activity of enhancers. Here the authors develop RamDA-seq, a method for full-length total RNA sequencing in single cells.
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影响因子：
5.8
作者：
Barnett, Derek W.;Garrison, Erik K.;Marth, Gabor T.
通讯作者：
Marth, Gabor T.
长链非编码 RNA 在癌症中的作用、功能和机制。
DOI：
10.1016/j.gpb.2015.09.006
发表时间：
2016-02
期刊：
Genomics, proteomics & bioinformatics
影响因子：
--
作者：
Fang Y;Fullwood MJ
通讯作者：
Fullwood MJ
影响因子：
46.9
作者：
Gaidatzis, Dimos;Burger, Lukas;Stadler, Michael B.
通讯作者：
Stadler, Michael B.
影响因子：
16.8
作者：
Ameur, Adam;Zaghlool, Ammar;Feuk, Lars
通讯作者：
Feuk, Lars
影响因子：
5.8
作者：
Angerer, Philipp;Haghverdi, Laleh;Buettner, Florian
通讯作者：
Buettner, Florian