Ab initio detection of multiple epitranscriptomic modifications from Oxford nanopore technology direct RNA sequencing...

Charting the eukaryotic epitranscriptome by direct RNA sequencing is promising but still very challenging, as current bioinformatics tools are based on modification-unaware software and require multiple modification-specific learning steps. Here, we introduce NanoSpeech, a modification-aware basecaller for the ab initio simultaneous detection of multiple modified bases using a transformer model, and NanoListener, which implements a simulated randomers strategy for robust training datasets and a new generation of ONT basecallers. NanoListener and NanoSpeech are independent of the specific ONT chemistry. Once a training dataset has been created, a single model with an expanded vocabulary can accurately basecall both unmodified and modified bases.