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Genome Editing in Penicillium chrysogenum Using Cas9 Ribonucleoprotein Particles

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Synthetic Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1772))

Abstract

Several CRISPR/Cas9 tools have been recently established for precise genome editing in a wide range of filamentous fungi. This genome editing platform offers high flexibility in target selection and the possibility of introducing genetic deletions without the introduction of transgenic sequences . This chapter describes an approach for the transformation of Penicillium chrysogenum protoplasts with preassembled ribonucleoprotein particles (RNPs) consisting of purified Cas9 protein and in vitro transcribed single guide RNA (sgRNA) for the deletion of genome sequences or their replacement with alternative sequences. This method is potentially transferable to all fungal strains where protoplasts can be obtained from.

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Acknowledgments

The work was supported by the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme FP7/2007-2013/under REA grant agreement no. [607332], and the Marie Skłodowska-Curie Co-funding of regional, national and international programmes (COFUND-DP) ALERT programme under REA grant agreement no. [713482].

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Authors and Affiliations

  1. Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands

    Carsten Pohl, László Mózsik, Arnold J. M. Driessen, Roel A. L. Bovenberg & Yvonne I. Nygård

  2. DSM Biotechnology Center, DSM Food Specialties B.V., Delft, The Netherlands

    Roel A. L. Bovenberg

  3. Chalmers University of Technology, Gothenburg, Sweden

    Yvonne I. Nygård

Authors
  1. Carsten Pohl
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  2. László Mózsik
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  3. Arnold J. M. Driessen
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  4. Roel A. L. Bovenberg
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  5. Yvonne I. Nygård
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Corresponding author

Correspondence to Arnold J. M. Driessen .

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Editors and Affiliations

  1. Agilent Technologies, Inc, La Jolla, California, USA

    Jeffrey Carl Braman

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Pohl, C., Mózsik, L., Driessen, A.J.M., Bovenberg, R.A.L., Nygård, Y. (2018). Genome Editing in Penicillium chrysogenum Using Cas9 Ribonucleoprotein Particles. In: Braman, J. (eds) Synthetic Biology. Methods in Molecular Biology, vol 1772. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7795-6_12

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  • DOI: https://doi.org/10.1007/978-1-4939-7795-6_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7794-9

  • Online ISBN: 978-1-4939-7795-6

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