Direct RNA sequencing enables m⁶A detection in endogenous transcript isoforms at base-specific resolution.

ABSTRACT: Direct RNA sequencing holds great promise for the de novo identification of RNA modifications at single-coordinate resolution; however, interpretation of raw sequencing output to discover modified bases remains a challenge. Using Oxford Nanopore's direct RNA sequencing technology, we developed a random forest classifier trained using experimentally detected N⁶-methyladenosine (m⁶A) sites within DRACH motifs. Our software MINES (m⁶A Identification using Nanopore Sequencing) assigned m⁶A methylation status to more than 13,000 previously unannotated DRACH sites in endogenous HEK293T transcripts and identified more than 40,000 sites with isoform-level resolution in a human mammary epithelial cell line. These sites displayed sensitivity to the

SUBMITTER: Lorenz DA 

PROVIDER: S-EPMC6913132 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature