Abstract
RNA modifications can influence gene expression via multiple aspects such as RNA stability and alternative splicing. The most prominent RNA modification is m6A (N6-methyladenosine). Its profiling from low starting amounts of <100 cells is challenging. We describe here a complete workflow from cell isolation to data analysis that is based on using an RNA CUT&RUN-supported m6A-RIP (RNA immunoprecipitation) procedure and a subsequent adaptor-tagging library synthesis. Male meiocytes isolated from maize anthers were used as a test system to establish the protocol.
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Acknowledgment
We thank the Meister lab for access to their MiSeq machine and excellent support by N. Eichner. This work was supported by the German Research Foundation DFG via Collaborative Research Center SFB960 to T.D. and a FAS grant (Financial Support for Equality) of the University of Regensburg to S.D..
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Shabani, D., Dresselhaus, T., Dukowic-Schulze, S. (2022). Profiling m6A RNA Modifications in Low Amounts of Plant Cells Using Maize Meiocytes. In: Lambing, C. (eds) Plant Gametogenesis. Methods in Molecular Biology, vol 2484. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2253-7_21
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DOI: https://doi.org/10.1007/978-1-0716-2253-7_21
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