Strawberry: Fast and accurate genome-guided transcript reconstruction and quantification from RNA-Seq

We propose a novel method and software tool, Strawberry, for transcript reconstruction and quantification from RNA-Seq data under the guidance of genome alignment and independent of gene annotation. Strawberry consists of two modules: assembly and quantification. The novelty of Strawberry is that the two modules use different optimization frameworks but utilize the same data graph structure, which allows a highly efficient, expandable and accurate algorithm for dealing large data. The assembly module parses aligned reads into splicing graphs, and uses network flow algorithms to select the most likely transcripts. The quantification module uses a latent class model to assign read counts from the nodes of splicing graphs to transcripts. Strawberry simultaneously estimates the transcript abundances and corrects for sequencing bias through an EM algorithm. Based on simulations, Strawberry outperforms Cufflinks and StringTie in terms of both assembly and quantification accuracies. Under the evaluation of a real data set, the estimated transcript expression by Strawberry has the highest correlation with Nanostring probe counts, an independent experiment measure for transcript expression. Availability: Strawberry is written in C++14, and is available as open source software at https://github.com/ruolin/strawberry under the MIT license.Author summary: Transcript assembly and quantification are important bioinformatics applications of RNA-Seq. The difficulty of solving these problem arises from the ambiguity of reads assignment to isoforms uniquely. This challenge is twofold: statistically, it requires a high-dimensional mixture model, and computationally, it needs to process datasets that commonly consist of tens of millions of reads. Existing algorithms either use very complex models that are too slow or assume no models, rather heuristic, and thus less accurate. Strawberry seeks to achieve a great balance between the model complexity and speed. Strawberry effectively leverages a graph-based algorithm to utilize all possible information from pair-end reads and, to our knowledge, is the first to apply a flow network algorithm on the constrained assembly problem. We are also the first to formulate the quantification problem in a latent class model. All of these features not only lead to a more flexible and complex quantification model but also yield software that is easier to maintain and extend. In this method paper, we have shown that the Strawberry method is novel, accurate, fast and scalable using both simulated data and real data.