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CRISPR/Cas9 Based Genome Editing of Penicillium chrysogenum

C. Pohl ; J. A. Kiel ; A. J. Driessen ; R. A. Bovenberg ; Yvonne Nygård (Institutionen för biologi och bioteknik, Industriell bioteknik)
ACS Synth Biol (2161-5063 (Electronic) 2161-5063 (Linking)). Vol. 5 (2016), 7, p. 754-64.
[Artikel, refereegranskad vetenskaplig]

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.

Nyckelord: Bacterial Proteins/genetics, *CRISPR-Cas Systems, DNA Repair, Endonucleases/genetics, Gene Deletion, Gene Editing/*methods, Gene Targeting/methods, Genetic Markers, Genetic Vectors, Genome, Fungal, Oligonucleotides/genetics, Penicillium chrysogenum/*genetics, RNA, Guide, *CRISPR/Cas9, *Penicillium chrysogenum, *Rnp, *genome editing, *marker-free gene deletion



Denna post skapades 2017-12-22. Senast ändrad 2018-01-17.
CPL Pubid: 254031

 

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Institutioner (Chalmers)

Institutionen för biologi och bioteknik, Industriell bioteknik

Ämnesområden

Genteknik inkl. funktionsgenomik

Chalmers infrastruktur