Large-scale RACE approach for proactive experimental definition of C. elegans ORFeome

Kourosh Salehi-Ashtiani, Chenwei Lin, Tong Hao, Yun Shen, David Szeto, Xinping Yang, Lila Ghamsari, Hanjoo Lee, Changyu Fan, Ryan R. Murray, Stuart Milstein, Nenad Svrzikapa, Michael E. Cusick, Frederick P. Roth, David E. Hill, Marc Vidal

Research output: Contribution to journalArticle

Abstract

Although a highly accurate sequence of the Caenorhabditis elegans genome has been available for 10 years, the exact transcript structures of many of its protein-coding genes remain unsettled. Approximately two-thirds of the ORFeome has been verified reactively by amplifying and cloning computationally predicted transcript models; still a full third of the ORFeome remains experimentally unverified. To fully identify the protein-coding potential of the worm genome including transcripts that may not satisfy existing heuristics for gene prediction, we developed a computational and experimental platform adapting rapid amplification of cDNA ends (RACE) for large-scale structural transcript annotation. We interrogated 2000 unverified protein-coding genes using this platform. We obtained RACE data for approximately two-thirds of the examined transcripts and reconstructed ORF and transcript models for close to 1000 of these. We defined untranslated regions, identified new exons, and redefined previously annotated exons. Our results show that as much as 20% of the C. elegans genome may be incorrectly annotated. Many annotation errors could be corrected proactively with our large-scale RACE platform.

Original languageEnglish (US)
Pages (from-to)2334-2342
Number of pages9
JournalGenome Research
Volume19
Issue number12
DOIs
StatePublished - Dec 2009

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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    Salehi-Ashtiani, K., Lin, C., Hao, T., Shen, Y., Szeto, D., Yang, X., Ghamsari, L., Lee, H., Fan, C., Murray, R. R., Milstein, S., Svrzikapa, N., Cusick, M. E., Roth, F. P., Hill, D. E., & Vidal, M. (2009). Large-scale RACE approach for proactive experimental definition of C. elegans ORFeome. Genome Research, 19(12), 2334-2342. https://doi.org/10.1101/gr.098640.109