A single-molecule long-read survey of the human transcriptome.
A single-molecule long-read survey of the human transcriptome.
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DOI：
10.1038/nbt.2705
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发表时间：
2013-11
影响因子：
46.9
通讯作者：
中科院分区：
文献类型：
--
作者：
全球RNA研究已成为理解生物过程的核心，但微阵列和短读测序等方法无法从5′端到3′端描述整个RNA分子。在这里，我们使用Pacific Biosciences的单分子长读测序技术对20个人体器官和组织的多聚腺苷酸化RNA互补序列进行测序，而无需片段化或扩增。我们表明，全长RNA分子高达1.5 kb，可以很容易地监测在5′端的序列丢失很少。对于较长的RNA分子，更多的5′核苷酸缺失，但完整的内含子结构通常被保留。我们总共鉴定了约14，000个剪接的GENCODE基因。高置信度映射与GENCODE注释一致，但>10%的比对表示先前未注释的内含子结构。作为一个群体，映射到未注释区域的转录本具有长的非编码RNA的特征。我们的研究结果显示了在单分子水平上对来自复杂真核转录组的全长RNA进行深度测序的可行性。
Global RNA studies have become central to understanding biological processes, but methods such as microarrays and short-read sequencing are unable to describe an entire RNA molecule from 5′ to 3′ end. Here we use single-molecule long-read sequencing technology from Pacific Biosciences to sequence the polyadenylated RNA complement of a pooled set of 20 human organs and tissues without the need for fragmentation or amplification. We show that full-length RNA molecules of up to 1.5 kb can readily be monitored with little sequence loss at the 5′ ends. For longer RNA molecules more 5′ nucleotides are missing, but complete intron structures are often preserved. In total, we identify ~14,000 spliced GENCODE genes. High-confidence mappings are consistent with GENCODE annotations, but >10% of the alignments represent intron structures that were not previously annotated. As a group, transcripts mapping to unannotated regions have features of long, noncoding RNAs. Our results show the feasibility of deep sequencing full-length RNA from complex eukaryotic transcriptomes on a single-molecule level.
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影响因子：
9.8
作者：
van Bakel H;Nislow C;Blencowe BJ;Hughes TR
通讯作者：
Hughes TR
影响因子：
4.5
作者：
Pickrell JK;Pai AA;Gilad Y;Pritchard JK
通讯作者：
Pritchard JK
影响因子：
12.3
作者：
通讯作者：
--
影响因子：
7
作者：
Harrow J;Frankish A;Gonzalez JM;Tapanari E;Diekhans M;Kokocinski F;Aken BL;Barrell D;Zadissa A;Searle S;Barnes I;Bignell A;Boychenko V;Hunt T;Kay M;Mukherjee G;Rajan J;Despacio-Reyes G;Saunders G;Steward C;Harte R;Lin M;Howald C;Tanzer A;Derrien T;Chrast J;Walters N;Balasubramanian S;Pei B;Tress M;Rodriguez JM;Ezkurdia I;van Baren J;Brent M;Haussler D;Kellis M;Valencia A;Reymond A;Gerstein M;Guigó R;Hubbard TJ
通讯作者：
Hubbard TJ
影响因子：
10.5
作者：
Cabili, Moran N.;Trapnell, Cole;Rinn, John L.
通讯作者：
Rinn, John L.