ORF Capture-Seq: a versatile method for targeted identification of full-length isoforms
| Main Authors: | Sheynkman, Gloria, Tuttle, Katharine, Laval, Florent, Tseng, Elizabeth, Underwood, Jason, Yu, Liang, Dong, Da, Smith, Melissa, Sebra, Robert, Willems, Luc, Hao, Tong, Calderwood, Michael, Hill, David, Vidal, Marc |
|---|---|
| Format: | Article eJournal |
| Bahasa: | eng |
| Terbitan: |
, 2020
|
| Subjects: | |
| Online Access: |
https://zenodo.org/record/3747665 |
Daftar Isi:
- Most human protein-coding genes are expressed as multiple isoforms. This in turn greatly expands the functional repertoire of the encoded proteome. While at least one reliable open reading frame (ORF) model has been assigned for every coding gene, the majority of alternative isoforms remains uncharacterized due to i) vast differences of overall levels between different isoforms expressed from common genes, and ii) the difficulty of obtaining full-length transcript sequences. Here, we present ORF Capture-Seq (OCS), a flexible method that addresses both challenges for targeted full-length isoform sequencing applications using collections of cloned ORFs as probes. As a proof-of-concept, we show that an OCS pipeline focused on genes coding for transcription factors increases isoform detection by an order of magnitude when compared to unenriched samples. In short, OCS enables rapid discovery of isoforms from custom-selected genes and will accelerate mapping of the human transcriptome.