SAGE2Splice: unmapped SAGE tags reveal novel splice junctions.

Serial analysis of gene expression (SAGE) not only is a method for profiling the global expression of genes, but also offers the opportunity for the discovery of novel transcripts. SAGE tags are mapped to known transcripts to determine the gene of origin. Tags that map neither to a known transcript...

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Main Authors: Byron Yu-Lin Kuo, Ying Chen, Slavita Bohacec, Ojvind Johansson, Wyeth W Wasserman, Elizabeth M Simpson
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2006-04-01
Series:PLoS Computational Biology
Online Access:https://journals.plos.org/ploscompbiol/article/file?id=10.1371/journal.pcbi.0020034&type=printable
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author Byron Yu-Lin Kuo
Ying Chen
Slavita Bohacec
Ojvind Johansson
Wyeth W Wasserman
Elizabeth M Simpson
author_facet Byron Yu-Lin Kuo
Ying Chen
Slavita Bohacec
Ojvind Johansson
Wyeth W Wasserman
Elizabeth M Simpson
author_sort Byron Yu-Lin Kuo
collection DOAJ
description Serial analysis of gene expression (SAGE) not only is a method for profiling the global expression of genes, but also offers the opportunity for the discovery of novel transcripts. SAGE tags are mapped to known transcripts to determine the gene of origin. Tags that map neither to a known transcript nor to the genome were hypothesized to span a splice junction, for which the exon combination or exon(s) are unknown. To test this hypothesis, we have developed an algorithm, SAGE2Splice, to efficiently map SAGE tags to potential splice junctions in a genome. The algorithm consists of three search levels. A scoring scheme was designed based on position weight matrices to assess the quality of candidates. Using optimized parameters for SAGE2Splice analysis and two sets of SAGE data, candidate junctions were discovered for 5%-6% of unmapped tags. Candidates were classified into three categories, reflecting the previous annotations of the putative splice junctions. Analysis of predicted tags extracted from EST sequences demonstrated that candidate junctions having the splice junction located closer to the center of the tags are more reliable. Nine of these 12 candidates were validated by RT-PCR and sequencing, and among these, four revealed previously uncharacterized exons. Thus, SAGE2Splice provides a new functionality for the identification of novel transcripts and exons. SAGE2Splice is available online at http://www.cisreg.ca.
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spelling doaj-art-1e37d59f178347f38c8e7805011bd15d2025-08-20T02:17:29ZengPublic Library of Science (PLoS)PLoS Computational Biology1553-734X1553-73582006-04-0124e3410.1371/journal.pcbi.0020034SAGE2Splice: unmapped SAGE tags reveal novel splice junctions.Byron Yu-Lin KuoYing ChenSlavita BohacecOjvind JohanssonWyeth W WassermanElizabeth M SimpsonSerial analysis of gene expression (SAGE) not only is a method for profiling the global expression of genes, but also offers the opportunity for the discovery of novel transcripts. SAGE tags are mapped to known transcripts to determine the gene of origin. Tags that map neither to a known transcript nor to the genome were hypothesized to span a splice junction, for which the exon combination or exon(s) are unknown. To test this hypothesis, we have developed an algorithm, SAGE2Splice, to efficiently map SAGE tags to potential splice junctions in a genome. The algorithm consists of three search levels. A scoring scheme was designed based on position weight matrices to assess the quality of candidates. Using optimized parameters for SAGE2Splice analysis and two sets of SAGE data, candidate junctions were discovered for 5%-6% of unmapped tags. Candidates were classified into three categories, reflecting the previous annotations of the putative splice junctions. Analysis of predicted tags extracted from EST sequences demonstrated that candidate junctions having the splice junction located closer to the center of the tags are more reliable. Nine of these 12 candidates were validated by RT-PCR and sequencing, and among these, four revealed previously uncharacterized exons. Thus, SAGE2Splice provides a new functionality for the identification of novel transcripts and exons. SAGE2Splice is available online at http://www.cisreg.ca.https://journals.plos.org/ploscompbiol/article/file?id=10.1371/journal.pcbi.0020034&type=printable
spellingShingle Byron Yu-Lin Kuo
Ying Chen
Slavita Bohacec
Ojvind Johansson
Wyeth W Wasserman
Elizabeth M Simpson
SAGE2Splice: unmapped SAGE tags reveal novel splice junctions.
PLoS Computational Biology
title SAGE2Splice: unmapped SAGE tags reveal novel splice junctions.
title_full SAGE2Splice: unmapped SAGE tags reveal novel splice junctions.
title_fullStr SAGE2Splice: unmapped SAGE tags reveal novel splice junctions.
title_full_unstemmed SAGE2Splice: unmapped SAGE tags reveal novel splice junctions.
title_short SAGE2Splice: unmapped SAGE tags reveal novel splice junctions.
title_sort sage2splice unmapped sage tags reveal novel splice junctions
url https://journals.plos.org/ploscompbiol/article/file?id=10.1371/journal.pcbi.0020034&type=printable
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AT slavitabohacec sage2spliceunmappedsagetagsrevealnovelsplicejunctions
AT ojvindjohansson sage2spliceunmappedsagetagsrevealnovelsplicejunctions
AT wyethwwasserman sage2spliceunmappedsagetagsrevealnovelsplicejunctions
AT elizabethmsimpson sage2spliceunmappedsagetagsrevealnovelsplicejunctions