| Autor: |
Wuestefeld T, Alexandre H. Thiery, Yue Wan, Ramanuj DasGupta, Sarah B. Ng, Lim Khe, Iakovleva, Chiea Chuen Khor, Toh Sy, Tran H, N. Davidson, Poon Syp, Fei Yao, Michael I. Love, Stanojevic D, Loo Jm, Mile Šikić, Li Chen, Ng Hev, Goh Wss, Watten L, Xuewen Ong, Koh Wqc, Yuk Kei Wan, Wee Joo Chng, Hwee Meng Low, Huck-Hui Ng, N.G. Iyer, Ng Hqa, Jia Xu Wang, Yu Q, Christopher Hendra, Harshil Patel, Sawyer C, Alicia Oshlack, Lee Pl, Sim A, J. Goeke, Tan Bop, Wai Leong Tam, Chan Y, L. Xin, Philip Ewels, Chen Y, Ploy N. Pratanwanich |
| Rok vydání: |
2021 |
| Předmět: |
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| DOI: |
10.1101/2021.04.21.440736 |
| Popis: |
The human genome contains more than 200,000 gene isoforms. However, different isoforms can be highly similar, and with an average length of 1.5kb remain difficult to study with short read sequencing. To systematically evaluate the ability to study the transcriptome at a resolution of individual isoforms we profiled 5 human cell lines with short read cDNA sequencing and Nanopore long read direct RNA, amplification-free direct cDNA, PCR-cDNA sequencing. The long read protocols showed a high level of consistency, with amplification-free RNA and cDNA sequencing being most similar. While short and long reads generated comparable gene expression estimates, they differed substantially for individual isoforms. We find that increased read length improves read-to-transcript assignment, identifies interactions between alternative promoters and splicing, enables the discovery of novel transcripts from repetitive regions, facilitates the quantification of full-length fusion isoforms and enables the simultaneous profiling of m6A RNA modifications when RNA is sequenced directly. Our study demonstrates the advantage of long read RNA sequencing and provides a comprehensive resource that will enable the development and benchmarking of computational methods for profiling complex transcriptional events at isoform-level resolution. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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