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Using CRISPR-Cas9 to Study ERK Signaling in Drosophila

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Book cover ERK Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1487))

Abstract

Genome engineering using the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated nuclease 9 (Cas9) technology is revolutionizing biomedical research. CRISPR-Cas9 enables precise editing of genes in a wide variety of cells and organisms, thereby accelerating molecular studies via targeted mutagenesis, epitope tagging, and other custom genetic modifications. Here, we illustrate the CRISPR-Cas9 methodology by focusing on Capicua (Cic), a nuclear transcriptional repressor directly phosphorylated and inactivated by ERK/MAPK. Specifically, we use CRISPR-Cas9 for targeting an ERK docking site of Drosophila Cic, thus generating ERK-insensitive mutants of this important signaling sensor.

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Acknowledgments

We thank A. Olza for Drosophila injections, N. Samper for experimental support, and F. Port, S. González-Crespo, Z. Paroush, M. Ruiz-Gómez, and A. Veraksa for discussions. This work was funded by grants from the Spanish Ministry of Science and Innovation (BFU2014-52863-P) and Fundació La Marató de TV3 (20131730). G.J. is an ICREA investigator.

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Authors and Affiliations

  1. Institut de Biologia Molecular de Barcelon-CSIC, Parc Cientific de Barcelona, Baldiri Reixac 10, 08028, Barcelona, Spain

    Marta Forés, Aikaterini Papagianni, Laura Rodríguez-Muñoz & Gerardo Jiménez

  2. ICREA, Pg. Lluís Companys 23, 08010, Barcelona, Spain

    Gerardo Jiménez

Authors
  1. Marta Forés
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  2. Aikaterini Papagianni
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  3. Laura Rodríguez-Muñoz
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  4. Gerardo Jiménez
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Corresponding author

Correspondence to Gerardo Jiménez .

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Editors and Affiliations

  1. Institut de Biologia Molecular de Barcelona-CSIC, Parc Científic de Barcelona, Barcelona, Spain

    Gerardo Jimenez

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Forés, M., Papagianni, A., Rodríguez-Muñoz, L., Jiménez, G. (2017). Using CRISPR-Cas9 to Study ERK Signaling in Drosophila . In: Jimenez, G. (eds) ERK Signaling. Methods in Molecular Biology, vol 1487. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6424-6_26

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  • DOI: https://doi.org/10.1007/978-1-4939-6424-6_26

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6422-2

  • Online ISBN: 978-1-4939-6424-6

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