CRISPR/Cas9 Based Genome Editing of Penicillium chrysogenum

• 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.

Subject headings

TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik -- Annan industriell bioteknik (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology -- Other Industrial Biotechnology (hsv//eng)

Keyword

Oligonucleotides/genetics

*genome editing

RNA

Genetic Markers

Fungal

Genome

*CRISPR-Cas Systems

*marker-free gene deletion

Penicillium chrysogenum/*genetics

Genetic Vectors

*Penicillium chrysogenum

Gene Editing/*methods

Gene Deletion

*Rnp

Bacterial Proteins/genetics

*CRISPR/Cas9

Gene Targeting/methods

Guide

Endonucleases/genetics

DNA Repair

Publication and Content Type

art (subject category)

ref (subject category)