The detecting of m6A RNA modifications upon Oxford Nanopore direct-RNA sequencing [Nanopore]

To investigate the potential mechanisms by which m6A contributes to ALS disease degeneration, we utilized a direct RNA sequencing platform, allowing for the identification of the m6A modification sites at single-nucleotide resolution. With the croreference of the ALS risk genes and transcriptomic RNA-seq data in ALS, we could further indntify the m6A-dependent potential genes and pathways in ALS. Overall design: We employed an optimized method using mouse ESC-derived motor neurons (MNs), matured with conditioned medium. This approach successfully generated MNs expressing mature neuronal markers and exhibiting longer, more developed neurite structures, as indicated by discrete Syn1-positive puncta. We then subjected these mature MNs to the ONT Nanopore platform, which leverages advanced machine-learning algorithms to detect RNA modifications through sequencing metrics.