@article{2a3b01d691d14f748fd05a43a496b1ed,
title = "Protocol for preparation and measurement of intracellular and extracellular modified RNA using liquid chromatography-mass spectrometry",
abstract = "About 150 modifications have been identified in RNA species. Besides their regulatory roles in the intracellular gene expression, abundant modified RNA nucleosides are catabolized from RNA and released into extracellular fluids, which can impact extracellular signaling as ligands for receptors. Here, we describe a protocol to prepare samples from biological specimens, including cultured cells, extracellular fluid, and tissues, to measure both intracellular and extracellular RNA modifications using mass spectrometry. For complete details on the use and execution of this protocol, please refer to Ogawa et al. (2021).",
keywords = "Mass Spectrometry, Metabolism, Metabolomics, Molecular Biology, Signal Transduction",
author = "Akiko Ogawa and Wei, {Fan Yan}",
note = "Funding Information: We are grateful for suggestions from all members of the Department of Molecular Physiology, Kumamoto University, and the Department of Modomics Biology & Medicine, Institute of Development, Aging and Cancer (IDAC), Tohoku University. We thank Y. Takahata and H. Miyamoto for technical assistance. This work was supported by JSPS KAKENHI grants 18H02599 (F.-Y.W.), 18K19521 (F.-Y.W.), 20H05309 (F.-Y.W.), and 20K18371 (A.O.), and by grants from the Japan Science and Technology Agency (JST), SAKIGAKE JPMJPR1532 (F.-Y.W.), Exploratory Research for Advanced Technology (ERATO) JPMJER2002 (F.-Y.W.), Fusion Oriented Research for disruptive Science and Technology (FOREST) JPMJFR205Y (F.-Y.W.), the Takeda Science Foundation (F.-Y.W.), the Uehara Memorial Foundation (F.-Y.W.), the Astellas Foundation for Research on Metabolic Disorders (A.O.), and the Japan Medical Women's Association Foundation (A.O.). A.O. performed experiments and analysis used in this study. F.-Y.W. developed the protocol. A.O. and F.-Y.W. wrote and revised this manuscript. The authors declare no competing interests. Funding Information: We are grateful for suggestions from all members of the Department of Molecular Physiology, Kumamoto University, and the Department of Modomics Biology & Medicine, Institute of Development, Aging and Cancer (IDAC), Tohoku University. We thank Y. Takahata and H. Miyamoto for technical assistance. This work was supported by JSPS KAKENHI grants 18H02599 (F.-Y.W.), 18K19521 (F.-Y.W.), 20H05309 (F.-Y.W.), and 20K18371 (A.O.), and by grants from the Japan Science and Technology Agency (JST), SAKIGAKE JPMJPR1532 (F.-Y.W.), Exploratory Research for Advanced Technology (ERATO) JPMJER2002 (F.-Y.W.), Fusion Oriented Research for disruptive Science and Technology (FOREST) JPMJFR205Y (F.-Y.W.), the Takeda Science Foundation (F.-Y.W.), the Uehara Memorial Foundation (F.-Y.W.), the Astellas Foundation for Research on Metabolic Disorders (A.O.), and the Japan Medical Women{\textquoteright}s Association Foundation (A.O.). Publisher Copyright: {\textcopyright} 2021 The Author(s)",
year = "2021",
month = dec,
day = "17",
doi = "10.1016/j.xpro.2021.100848",
language = "English",
volume = "2",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "4",
}