SAGETTARIUS: a program to reduce the number of tags mapped to multiple transcripts and to plan SAGE sequencing stages

Abstract

SAGE (Serial Analysis of Gene Expression) experiments generate short nucleotide sequences called 'tags' which are assumed to map unambiguously to their original transcripts (1 tag to 1 transcript mapping). Nevertheless, many tags are generated that do not map to any transcript or map to multiple transcripts. Current bioinformatics resources, such as SAGEmap and TAGmapper, have focused on reducing the number of unmapped tags. Here, we describe SAGETTARIUS, a new high-throughput program that performs successive precise Nla3 and Sau3A tag to transcript mapping, based on specifically designed Virtual Tag (VT) libraries. First, SAGETTARIUS decreases the number of tags mapped to multiple transcripts. Among the various mapping resources compared, SAGETTARIUS performed the best in this respect by decreasing up to 11% the number of multiply mapped tags. Second, SAGETTARIUS allows the establishment of a guideline for SAGE experiment sequencing efforts through efficient mapping of the CRT (Cytoplasmic Ribosomal protein Transcripts)-specific tags. Using all publicly available human and mouse Nla3 SAGE experiments, we show that sequencing 100,000 tags is sufficient to map almost all CRT-specific tags and that four sequencing stages can be identified when carrying out a human or mouse SAGE project. SAGETTARIUS is web interfaced and freely accessible to academic users.

Figure 1.

Generation of SAGETTARIUS database information. The final results of the procedure are VT to transcript associations. Bold: procedure steps.

Figure 2.

SAGETTARIUS progressive and reductive ET to transcript mapping process.

Figure 3.

Progressive detection of CRT-specific tags in (a) 371 human Nla3 SAGE experiments with the number of sequenced ET ranging from 1430 to 308 589 and (b) 123 mouse Nla3 SAGE experiments with the number of sequenced ET ranging from 464 to 194 345. In both human and mouse, SAGE experiments can be divided into four major sequencing stages (- -) based on the detection of CRT-specific tags.

Figure 4.

Number of unmapped, multiply mapped and single-mapped ET sequences from the GSM14740 SAGE experiment (40 027 unique ET sequences). ET mappings have been carried out by SAGETTARIUS, TAGmapper, SAGEmap-reliable and SAGEmap-full resources.