RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome

RSEM is a software package for estimating gene and isoform expression levels from RNA-Seq data. The new RSEM package (rsem-1.x) provides an user-friendly interface, supports threads for parallel computation of the EM algorithm, single-end and paired-end read data, quality scores, variable-length reads and RSPD estimation. It can also generate genomic-coordinate BAM files and UCSC wiggle files for visualization. In addition, it provides posterior mean and 95% credibility interval estimates for expression levels.

A key challenge in transcript quantification from RNA-Seq data is the handling of reads that map to multiple genes or isoforms. This issue is particularly important for quantification with de novo transcriptome assemblies in the absence of sequenced genomes, as it is difficult to determine which transcripts are isoforms of the same gene. A second significant issue is the design of RNA-Seq experiments, in terms of the number of reads, read length, and whether reads come from one or both ends of cDNA fragments.

We have presented RSEM, a software package for performing gene and isoform level quantification from RNA-Seq data. Through simulations and evaluations with real data, we have shown that RSEM has superior or comparable performance to other quantification methods. Unlike other tools, RSEM does not require a reference genome and thus should be useful for quantification with de novo transcriptome assemblies. The software package has a number of other useful features for RNA-Seq researchers including visualization outputs and CI estimates. In addition, the software is user-friendly, typically requiring at most two commands to estimate abundances from raw RNA-Seq reads and uses reference transcript files in standard formats. Lastly, RSEM’s simulation module is valuable for determining optimal sequencing strategies for quantification experiments. Taking advantage of this module, we have determined that a large number of short SE reads is best for gene-level quantification, while PE reads may improve within-gene isoform frequencies for the mouse and human transcript sets.

Li B, Dewey CN. (2011) RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinformatics 12, 323. [article]

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