Abstract
In highly polarized cells such as neurons, most RNA molecules are not randomly distributed but sorted into different compartments. So far, methods to analyze the transcriptome in distinct subcellular compartments are not well established. Here, we first describe the culturing of primary motoneurons in compartmentalized chambers to separate the axons from the somatodendritic compartment. Second, we introduce a method for whole transcriptome amplification followed by high-throughput sequencing to analyze the RNA composition of these two different compartments in neuronal cells.
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Acknowledgments
This work was supported by Deutsche Forschungsgemeinschaft (BR4910/1-1 to M.B., SE697/4-1 to M.S.); Deutsche Forschungsgemeinschaft (BR4910/2-1 to M.B., SE697/5-1 to M.S.); Bayerischer Forschungsverbund Induzierte Pluripotente Stammzellen (D2-F2412.26 to M.S.); European Community’s Health Seventh Framework Programme through the Euro-MOTOR Consortium (259867 to M.S.); and the Hermann und Lilly Schilling–Stiftung im Stifterverband der Deutschen Industrie.
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Saal-Bauernschubert, L., Briese, M., Sendtner, M. (2017). Optimized Whole Transcriptome Profiling of Motor Axons. In: Kaufmann, M., Klinger, C., Savelsbergh, A. (eds) Functional Genomics. Methods in Molecular Biology, vol 1654. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7231-9_16
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DOI: https://doi.org/10.1007/978-1-4939-7231-9_16
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7230-2
Online ISBN: 978-1-4939-7231-9
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