Targeted RNA sequencing reveals the deep complexity of the human transcriptome.

Targeted RNA sequencing reveals the deep complexity of the human transcriptome.
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DOI:
10.1038/nbt.2024
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发表时间:
2011-11-13
影响因子:
46.9
通讯作者:
--
中科院分区:
工程技术1区
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--
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转录组学分析揭示了人类转录组的意想不到的复杂性,其广度和深度超过了目前的RNA测序能力。使用平铺阵列的目标和序列选择部分的转录组,我们确定和表征未注释的转录本,其罕见的或短暂的表达低于常规测序方法的检测限。我们使用这种技术提供的前所未有的覆盖深度来达到人类转录组的最深极限,暴露基因间区域中广泛的,受调控的和非常复杂的非编码转录,以及甚至深入研究的蛋白质编码位点(如p53和HOX)中的未注释外显子和剪接模式。数据还显示,在常规RNA测序数据集中观察到的间歇性测序读数(先前被视为噪音)实际上指示未组装的罕见转录物。总的来说,这些结果揭示了人类转录组的范围,深度和复杂性,这远远没有得到充分的表征。
Transcriptomic analyses have revealed an unexpected complexity to the human transcriptome, whose breadth and depth exceeds current RNA sequencing capability. Using tiling arrays to target and sequence select portions of the transcriptome, we identify and characterize unannotated transcripts whose rare or transient expression is below the detection limits of conventional sequencing approaches. We use the unprecedented depth of coverage afforded by this technique to reach the deepest limits of the human transcriptome, exposing widespread, regulated and remarkably complex noncoding transcription in intergenic regions, as well as unannotated exons and splicing patterns in even intensively studied protein-coding loci such as p53 and HOX. The data also show that intermittent sequenced reads observed in conventional RNA sequencing data sets, previously dismissed as noise, are in fact indicative of unassembled rare transcripts. Collectively, these results reveal the range, depth and complexity of a human transcriptome that is far from fully characterized.
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