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Characterizing RNA modifications in the central nervous system and single cells by RNA sequencing and liquid chromatography–tandem mass spectrometry techniques

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Abstract

Post-transcriptional modifications to RNA constitute a newly appreciated layer of translation regulation in the central nervous system (CNS). The identity, stoichiometric quantity, and sequence position of these unusual epitranscriptomic marks are central to their function, making analytical methods that are capable of accurate and reproducible measurements paramount to the characterization of the neuro-epitranscriptome. RNA sequencing–based methods and liquid chromatography–tandem mass spectrometry (LC–MS/MS) techniques have been leveraged to provide an early glimpse of the landscape of RNA modifications in bulk CNS tissues. However, recent advances in sample preparation, separations, and detection methods have revealed that individual cells display remarkable heterogeneity in their RNA modification profiles, raising questions about the prevalence and function of cell-specific distributions of post-transcriptionally modified nucleosides in the brain. In this Trends article, we present an overview of RNA sequencing and LC–MS/MS methodologies for the analysis of RNA modifications in the CNS with special emphasis on recent advancements in techniques that facilitate single-cell and subcellular detection.

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  1. Department of Chemistry, Tufts University, Medford, MA, 02155, USA

    Arya Patel & Kevin D. Clark

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Published in the topical collection Young Investigators in (Bio-)Analytical Chemistry 2023 with guest editors Zhi-Yuan Gu, Beatriz Jurado-Sánchez, Thomas H. Linz, Leandro Wang Hantao, Nongnoot Wongkaew, and Peng Wu.

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Patel, A., Clark, K.D. Characterizing RNA modifications in the central nervous system and single cells by RNA sequencing and liquid chromatography–tandem mass spectrometry techniques. Anal Bioanal Chem 415, 3739–3748 (2023). https://doi.org/10.1007/s00216-023-04604-y

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