Genome Editing in Penicillium chrysogenum Using Cas9 Ribonucleoprotein Particles

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

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

13 Citations (Scopus)
84 Downloads (Pure)


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.

Original languageEnglish
Title of host publicationSynthetic biology
Subtitle of host publicationMethods and Protocols
EditorsJeffrey Carl Braman
Place of PublicationNew York
PublisherHumana Press
Number of pages20
ISBN (Electronic)978-1-4939-7795-6
ISBN (Print)978-1-4939-7794-9
Publication statusPublished - May-2018

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


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

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