Common analysis of direct RNA sequencinG CUrrently leads to misidentification of m⁵C at GCU motifs.

ABSTRACT: RNA modifications, such as methylation, can be detected with Oxford Nanopore Technologies direct RNA sequencing. One commonly used tool for detecting 5-methylcytosine (m⁵C) modifications is Tombo, which uses an "Alternative Model" to detect putative modifications from a single sample. We examined direct RNA sequencing data from diverse taxa including viruses, bacteria, fungi, and animals. The algorithm consistently identified a m⁵C at the central position of a GCU motif. However, it also identified a m⁵C in the same motif in fully unmodified in vitro transcribed RNA, suggesting that this is a frequent false prediction. In the absence of further validation, several published predictions of m⁵C in a GCU context should be reconsidered, including tho

SUBMITTER: Watson KJ 

PROVIDER: S-EPMC10687253 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature