A fast and globally optimal solution for RNA-seq quantification.

Alignment-based RNA-seq quantification methods typically involve a time-consuming alignment process prior to estimating transcript abundances. In contrast, alignment-free RNA-seq quantification methods bypass this step, resulting in significant speed improvements. Existing alignment-free methods rely on the Expectation-Maximization (EM) algorithm for estimating transcript abundances. However, EM algorithms only guarantee locally optimal solutions, leaving room for further accuracy improvement by finding a globally optimal solution. In this study, we present TQSLE, the first alignment-free RNA-seq quantification method that provides a globally optimal solution for transcript abundances estimation. TQSLE adopts a two-step approach: first, it constructs a k-mer frequency matrix A for the reference transcriptome and a k-mer frequency vector b for the RNA-seq reads; then, it directly estimates transcript abundances by solving the linear equation ATAx = ATb. We evaluated the performance of TQSLE using simulated and real RNA-seq data sets and observed that, despite comparable speed to other alignment-free methods, TQSLE outperforms them in terms of accuracy. TQSLE is freely available at https://github.com/yhg926/TQSLE.