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Genome Editing: CRISPR-Cas9

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Book cover Fungal Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1775))

Abstract

In the present chapter, we present the protocols and guidelines to facilitate implementation of CRISPR-Cas9 technology in fungi where few or no genetic tools are in place. Hence, we firstly explain how to identify dominant markers for genetic transformation. Secondly, we provide a guide for construction of Cas9/sgRNA episomal expression vectors. Thirdly, we present how to mutagenize reporter genes to explore the efficiency of CRISPR-Cas9 in the relevant fungus and to ease subsequent CRISPR-mediated genetic engineering. Lastly, we describe how to make CRISPR-mediated marker-dependent and marker-free gene targeting.

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Author information

Authors and Affiliations

  1. Department of Biotechnology and Biomedicine, Technical University of Denmark, Kgs.Lyngby, Denmark

    Jakob B. Hoof, Christina S. Nødvig & Uffe H. Mortensen

Authors
  1. Jakob B. Hoof
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  2. Christina S. Nødvig
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  3. Uffe H. Mortensen
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Corresponding author

Correspondence to Uffe H. Mortensen .

Editor information

Editors and Affiliations

  1. Fungal Physiology, Westerdijk Fungal, Biodiversity Institute, Utrecht, The Netherlands

    Ronald P. de Vries

  2. Centre for Structural and Functional Genomics, Concordia University, Montreal, Québec, Canada

    Adrian Tsang

  3. US Department of Energy, Joint Genome Institute, Walnut Creek, California, USA

    Igor V. Grigoriev

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Hoof, J.B., Nødvig, C.S., Mortensen, U.H. (2018). Genome Editing: CRISPR-Cas9. In: de Vries, R., Tsang, A., Grigoriev, I. (eds) Fungal Genomics. Methods in Molecular Biology, vol 1775. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7804-5_11

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  • DOI: https://doi.org/10.1007/978-1-4939-7804-5_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7803-8

  • Online ISBN: 978-1-4939-7804-5

  • eBook Packages: Springer Protocols

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