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All the sites we cannot see: Sources and mitigation of false negatives in RNA modification studies. | LitMetric

All the sites we cannot see: Sources and mitigation of false negatives in RNA modification studies.

Nat Rev Mol Cell Biol

Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.

Published: October 2024

AI Article Synopsis

Article Abstract

RNA modifications are essential for human health - too much or too little of them leads to serious illnesses ranging from neurodevelopmental disorders to cancer. Technical advances in RNA modification sequencing are beginning to uncover the RNA targets of diverse RNA-modifying enzymes that are dysregulated in disease. However, the emerging transcriptome-wide maps of modified nucleosides installed by these enzymes should be considered as first drafts. In particular, a range of technical artefacts lead to false negatives - modified sites that are overlooked owing to technique-dependent, and often sequence-context-specific, 'blind spots'. In this Review, we discuss potential sources of false negatives in sequencing-based RNA modification maps, propose mitigation strategies and suggest guidelines for transparent reporting of sensitivity to detect modified sites in profiling studies. Important considerations for recognition and avoidance of false negatives include assessment and reporting of position-specific sequencing depth, identification of protocol-dependent RNA capture biases and applying controls for false negatives as well as for false positives. Despite their limitations, emerging maps of RNA modifications reveal exciting and largely uncharted potential for post-transcriptional control of all aspects of RNA function.

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http://dx.doi.org/10.1038/s41580-024-00784-2DOI Listing

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