CRISPR/Cas9 Based Genome Editing of Penicillium chrysogenum
Journal article, 2016

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.


*genome editing


Genetic Markers



*CRISPR-Cas Systems

*marker-free gene deletion

Penicillium chrysogenum/*genetics

Genetic Vectors

*Penicillium chrysogenum

Gene Editing/*methods

Gene Deletion


Bacterial Proteins/genetics


Gene Targeting/methods



DNA Repair


C. Pohl

University of Groningen

J. A. Kiel

University of Groningen

A. J. Driessen

University of Groningen

R. A. Bovenberg

DSM Biotechnology Centre

University of Groningen

Yvonne Nygård

University of Groningen

ACS Synthetic Biology

2161-5063 (eISSN)

Vol. 5 7 754-64

Subject Categories

Other Industrial Biotechnology



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