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Detection of Nascent RNA Transcripts by Fluorescence In Situ Hybridization

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Fluorescence in situ Hybridization (FISH)

Part of the book series: Methods in Molecular Biology ((MIMB,volume 659))

Abstract

The development of cellular diversity within any organism depends on the timely and correct expression of differing subsets of genes within each tissue type. Many techniques exist which allow a global, average analysis of RNA expression; however, RNA-FISH permits the sensitive detection of specific transcripts within individual cells while preserving the cellular morphology. The technique can provide insight into the spatial and temporal organization of gene transcription as well the relationship of gene expression and mature RNA distribution to nuclear and cellular compartments. It can also reveal the intercellular variation of gene expression within a given tissue. Here, we describe RNA-FISH methodologies that allow the detection of nascent transcripts within the cell nucleus as well as protocols that allow the detection of RNA alongside DNA or proteins. Such techniques allow the placing of gene transcription within a functional context of the whole cell.

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Acknowledgments

We thank the Grosveld Lab in Rotterdam, particularly Peter Fraser, Joost Gribnau, Tolleiv Trimborn, and Mark Wijgerde for initial support with setting up RNA-FISH as a technique in our laboratory. We thank Jackie Sloane-Stanley and Sue Butler for provision of cells. Our work is funded by the Medical Research Council.

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Authors and Affiliations

  1. MRC Molecular Haematology Unit, John Radcliffe Hospital, Weatherall Institute of Molecular Medicine, Headington, Oxford, UK

    Jill M. Brown & Veronica J. Buckle

Authors
  1. Jill M. Brown
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  2. Veronica J. Buckle
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Corresponding author

Correspondence to Jill M. Brown .

Editor information

Editors and Affiliations

  1. School of Health Sciences & Social Care, Div. Biosciences, Brunel University, Kingston Lane, Uxbridge, UB83PH, United Kingdom

    Joanna M. Bridger

  2. , Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX37BN, United Kingdom

    Emanuela V. Volpi

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Brown, J.M., Buckle, V.J. (2010). Detection of Nascent RNA Transcripts by Fluorescence In Situ Hybridization. In: Bridger, J., Volpi, E. (eds) Fluorescence in situ Hybridization (FISH). Methods in Molecular Biology, vol 659. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-789-1_3

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  • DOI: https://doi.org/10.1007/978-1-60761-789-1_3

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-788-4

  • Online ISBN: 978-1-60761-789-1

  • eBook Packages: Springer Protocols

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