Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia.
Nat Biotechnol. 2011 Nov 13;30(1):99-104. doi: 10.1038/nbt.2024.
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.
转录组分析揭示了人类转录组出人意料的复杂性，其广度和深度超过了当前 RNA 测序的能力。我们使用平铺阵列来靶向和测序转录组的选定部分，从而鉴定和描述那些稀有或瞬时表达低于传统测序方法检测限的未注释转录本。我们利用这项技术前所未有的深度覆盖范围，达到了人类转录组的最深极限，揭示了广泛存在的、受调控的、非常复杂的非编码转录，以及在 p53 和 HOX 等即使经过深入研究的蛋白编码基因座中未注释的外显子和剪接模式。这些数据还表明，在常规 RNA 测序数据集中观察到的间歇性测序读段，以前被认为是噪声，实际上表明了未组装的稀有转录本。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，远远没有得到充分的描述。
