Publication

CRISPR/Cas9 based genome editing of Penicillium chrysogenum

Pohl, C., Kiel, J. A. K. W., Driessen, A. J. M., Bovenberg, R. A. L. & Nygård, Y., 12-Apr-2016, In : ACS Synthetic Biology. 5, 7, p. 754-764 11 p., 6b00082.

Research output: Contribution to journalArticleAcademicpeer-review

Copy link to clipboard

Documents

  • CRISPR/Cas9 Based Genome Editing of Penicillium chrysogenum

    Final publisher's version, 497 KB, PDF-document

    Request copy

DOI

CRISPR/Cas9 based systems have emerged as versatile platforms for precision genome editing in a wide range of organisms. Here we have developed powerful CRISPR/Cas9 tools for marker-based and marker-free genome modifications in Penicillium chrysogenum, a model filamentous fungus and industrially relevant cell factory. The developed CRISPR/Cas9 toolbox is highly flexible and allows editing of new targets with minimal cloning efforts. The Cas9 protein and the sgRNA can be either delivered during transformation, as preassembled CRISPR-Cas9 ribonucleoproteins (RNPs) or expressed from an AMA1 based plasmid within the cell. The direct delivery of the Cas9 protein with in vitro synthesized sgRNA to the cells allows for a transient method for genome engineering that may rapidly be applicable for other filamentous fungi. The expression of Cas9 from an AMA1 based vector was shown to be highly efficient for marker-free gene deletions.

Original languageEnglish
Article number6b00082
Pages (from-to)754-764
Number of pages11
JournalACS Synthetic Biology
Volume5
Issue number7
Publication statusPublished - 12-Apr-2016

    Keywords

  • CRISPR/Cas9, genome editing, marker-free gene deletion, Penicillium chrysogenum, RNP, FUNGUS, MUTAGENESIS, ASPERGILLUS-FUMIGATUS, MEDIATED DELIVERY, GENE DISRUPTION, HUMAN-CELLS, GUIDE RNA, CAS9, RIBONUCLEOPROTEINS, SYSTEM

View graph of relations

ID: 31541424