Background: The accurate characterization of RNA transcripts and expression levels across species is critical for understanding transcriptome evolution. As available RNA-seq data accumulate rapidly, there is a great demand for tools that build gene annotations for cross-species RNA-seq analysis. However, prevailing methods of ortholog annotation for RNA-seq analysis between closely-related species do not take inter-species variation in mappability into consideration.
Results: Here we present XSAnno, a computational framework that integrates previous approaches with multiple filters to improve the accuracy of inter-species transcriptome comparisons. The implementation of this approach in comparing RNA-seq data of human, chimpanzee, and rhesus macaque brain transcriptomes has reduced the false discovery of differentially expressed genes, while maintaining a low false negative rate.
Conclusion: The present study demonstrates the utility of the XSAnno pipeline in building ortholog annotations and improving the accuracy of cross-species transcriptome comparisons.