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In Embryo Gene Reporter Assays for Evaluation of Cis-Regulatory Regions

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DNA-Protein Interactions

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

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Abstract

Gene expression reporter assays measure the relevance of cis-regulatory elements and DNA-binding proteins in modulating transcriptional activity. Commonly, they are performed in cell lines. However, regulation of transcriptional activity during development is complex and dynamic, and not many cell lines reproduce the embryonic conditions. Thus, conclusions derived from cell line data provide limited information about embryonic development. On the other hand, one of the major hurdles for embryonic assays is delivering reporter plasmids in a tissue-specific manner. In this sense, the chick embryo is a good model system to perform these assays. Electroporation of chick embryos provides temporal and spatially controlled plasmid delivery. Further, it is a well-established, easy, and an economical procedure. Here, we describe in detail how to measure in the chick neural tube (1) enhancer activity with GFP, (2) enhancer activity with luciferase, and (3) 3′UTR activity with luciferase.

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Author information

Authors and Affiliations

  1. Department of Cell and Developmental Biology, Biomedical Sciences Institute, Universidade de São Paulo, São Paulo, Brazil

    Carolina Purcell Goes & C. Y. Irene Yan

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

    Tatiane Y. Kanno

Authors
  1. Carolina Purcell Goes
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  2. Tatiane Y. Kanno
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  3. C. Y. Irene Yan
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Corresponding author

Correspondence to C. Y. Irene Yan .

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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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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Goes, C.P., Kanno, T.Y., Yan, C.Y.I. (2023). In Embryo Gene Reporter Assays for Evaluation of Cis-Regulatory Regions. 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_16

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  • DOI: https://doi.org/10.1007/978-1-0716-2847-8_16

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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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