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In Situ Hybridization as a Method to Examine Gene Regulatory Activity In Vivo

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DNA-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2599))

Abstract

Transcription factor-enhancer binding events are among the most well-studied protein-DNA interactions, allowing researchers to determine mechanisms of transcriptional activation or repression during development. While large-scale ChIP-sequence datasets, together with computational predictions and chromatin accessibility data, yield information on potential transcription factor binding activities, reporter gene assays provide measurable information on whether these binding activities are functional in particular cell types during development. Here, we present a detailed protocol to examine enhancer activity in Drosophila embryos using cloning, transgenesis, and in situ hybridization.

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Author information

Authors and Affiliations

  1. Department of Biology, Hamilton College, Clinton, NY, USA

    Rhea R. Datta

  2. Department of Molecular Biosciences, Northwestern University, Evanston, IL, USA

    Pinar Onal

  3. Department of Molecular Biology and Genetics, Ihsan Dogramaci Bilkent University, Ankara, Turkey

    Pinar Onal

Authors
  1. Rhea R. Datta
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  2. Pinar Onal
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Corresponding authors

Correspondence to Rhea R. Datta or Pinar Onal .

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Editors and Affiliations

  1. Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA

    Marcos Simoes-Costa

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Datta, R.R., Onal, P. (2023). In Situ Hybridization as a Method to Examine Gene Regulatory Activity In Vivo. In: Simoes-Costa, M. (eds) DNA-Protein Interactions. Methods in Molecular Biology, vol 2599. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2847-8_17

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  • DOI: https://doi.org/10.1007/978-1-0716-2847-8_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2846-1

  • Online ISBN: 978-1-0716-2847-8

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