Simultaneous isoform discovery and quantification from RNA-seq

RNA sequencing is a recent technology which has seen an explosion of methods addressing all levels of analysis, from read mapping to transcript assembly to differential expression modeling. In particular the discovery of isoforms at the transcript assembly stage is a complex problem and current approaches suffer from various limitations. For instance, many approaches use graphs to construct a minimal set of isoforms which covers the observed reads, then perform a separate algorithm to quantify the isoforms, which can result in a loss of power. Current methods also use ad-hoc solutions to deal with the vast number of possible isoforms which can be constructed from a given set of reads. Finally, while the need of taking into account features such as read pairing and sampling rate of reads has been acknowledged, most existing methods do not seamlessly integrate these features as part of the model. We present Montebello, an integrated statistical approach which performs simultaneous isoform discovery and quantification by using a Monte Carlo simulation to find the most likely isoform composition leading to a set of observed reads. We compare Montebello to Cufflinks, a popular isoform discovery approach, on a simulated data set and on 46.3 million brain reads from an Illumina tissue panel. On this data set Montebello appears to offer a modest improvement over Cufflinks when considering discovery and parsimony metrics. In addition Montebello mitigates specific difficulties inherent in the Cufflinks approach. Finally, Montebello can be fine-tuned depending on the type of solution desired.