Genome editing in penicillium chrysogenum using cas9 ribonucleoprotein particles

Carsten Pohl, László Mózsik, Arnold J.M. Driessen (Corresponding author), Roel A.L. Bovenberg, Yvonne I. Nygård

Research output: Chapter in Book/Report/Conference proceedingChapter or book articleScientificpeer-review

18 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationSynthetic Biology: Methods and Protocols
PublisherHumana Press
Pages213-232
Number of pages20
ISBN (Electronic)978-1-4939-7795-6
ISBN (Print)978-1-4939-7794-9, 978-1-4939-9287-4
DOIs
Publication statusPublished - 2018
MoE publication typeA3 Part of a book or another research book

Publication series

SeriesMethods in Molecular Biology
Volume1772
ISSN1064-3745

Keywords

  • CRISPR/Cas9
  • Filamentous fungi
  • Genome editing
  • HDR
  • NHEJ
  • Penicillium chrysogenum
  • Ribonucleoprotein particle
  • RNP
  • Endonucleases/genetics
  • Protoplasts/metabolism
  • Gene Targeting/methods
  • Gene Editing/methods
  • RNA, Guide, CRISPR-Cas Systems/genetics
  • CRISPR-Cas Systems/genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics
  • Gene Deletion
  • Ribonucleoproteins/genetics
  • Penicillium chrysogenum/genetics

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