Detection of m6A from direct RNA sequencing using a multiple instance learning framework

RNA modifications such as m6A methylation form an additional layer of complexity in the transcriptome. Nanopore direct RNA sequencing can capture this information in the raw current signal for each RNA molecule, enabling the detection of RNA modifications using supervised machine learning. However, experimental approaches provide only site-level training data, whereas the modification status for each single RNA molecule is missing. Here we present m6Anet, a neural-network-based method that leverages the multiple instance learning framework to specifically handle missing read-level modification labels in site-level training data. m6Anet outperforms existing computational methods, shows similar accuracy as experimental approaches, and generalizes with high accuracy to different cell lines and species without retraining model parameters. In addition, we demonstrate that m6Anet captures the underlying read-level stoichiometry, which can be used to approximate differences in modification rates. Overall, m6Anet offers a tool to capture the transcriptome-wide identification and quantification of m6A from a single run of direct RNA sequencing. This work presents m6Anet, which implements a neural-network-based multiple instance learning model to detect m6A modifications from direct RNA sequencing data.

Automated summary

This study introduces a neural-network-based method m6Anet, which leverages multiple instance learning framework to handle missing read-level modification labels in site-level training data, demonstrating superior performance in detecting m6A modifications.