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Live Imaging of Nuclear RNPs in Mammalian Complex Tissue with ECHO-liveFISH

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RNA Detection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1649))

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Abstract

Multiplex RNA detection with fluorescence microscopy offers high spatial and temporal resolution required for addressing complex behaviors of RNA in living cells. Using chemically engineered linear oligonucleotide probes that emit fluorescence upon hybridization to target RNA, we have devised an imaging method suitable for studies of the dynamic regulation of nuclear RNPs, an important and yet poorly understood cellular pathway of gene expression. This new method labels specific sequences of RNA components in RNPs and thus avoids overexpression of fluorescent marker proteins that may result in entangled experimental results. Using this method, we observe in living brain tissue spatially constrained nuclear RNA foci under dynamic regulation in response to cellular transcriptional activity with individual cell heterogeneity.

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Author information

Authors and Affiliations

  1. Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-Honmachi, Sakyo-ku, Kyoto, 606-8501, Japan

    Dan Ohtan Wang

  2. The Keihanshin Consortium for Fostering the Next Generation of Global Leaders in Research (K-CONNEX), Yoshida-Honmachi, Sakyo-ku, Kyoto, 606-8302, Japan

    Dan Ohtan Wang

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  1. Dan Ohtan Wang
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Correspondence to Dan Ohtan Wang .

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Editors and Affiliations

  1. Developmental Biology Unit, EMBL Heidelberg, Heidelberg, Germany

    Imre Gaspar

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Wang, D.O. (2018). Live Imaging of Nuclear RNPs in Mammalian Complex Tissue with ECHO-liveFISH. In: Gaspar, I. (eds) RNA Detection. Methods in Molecular Biology, vol 1649. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7213-5_17

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  • DOI: https://doi.org/10.1007/978-1-4939-7213-5_17

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7212-8

  • Online ISBN: 978-1-4939-7213-5

  • eBook Packages: Springer Protocols

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