Genome Editing in Penicillium chrysogenum Using Cas9 Ribonucleoprotein Particles

  • Carsten Pohl
  • László Mózsik
  • Arnold J. M. DriessenEmail author
  • Roel A. L. Bovenberg
  • Yvonne I. Nygård
Part of the Methods in Molecular Biology book series (MIMB, volume 1772)


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.


Ribonucleoprotein particle RNP CRISPR/Cas9 Genome editing NHEJ HDR Penicillium chrysogenum Filamentous fungi 



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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Carsten Pohl
    • 1
  • László Mózsik
    • 1
  • Arnold J. M. Driessen
    • 1
    Email author
  • Roel A. L. Bovenberg
    • 1
    • 2
  • Yvonne I. Nygård
    • 1
    • 3
  1. 1.Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology InstituteUniversity of GroningenGroningenThe Netherlands
  2. 2.DSM Biotechnology CenterDSM Food Specialties B.V.DelftThe Netherlands
  3. 3.Chalmers University of TechnologyGothenburgSweden

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