CRISPR/Cas9 Based Genome Editing of Penicillium chrysogenum
Artikel i vetenskaplig tidskrift, 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.

*Penicillium chrysogenum

Guide

Oligonucleotides/genetics

Fungal

*CRISPR/Cas9

Genetic Vectors

Penicillium chrysogenum/*genetics

RNA

*Rnp

Bacterial Proteins/genetics

Genetic Markers

*marker-free gene deletion

Genome

*genome editing

Gene Deletion

DNA Repair

Gene Editing/*methods

Endonucleases/genetics

*CRISPR-Cas Systems

Gene Targeting/methods

Författare

C. Pohl

J. A. Kiel

A. J. Driessen

R. A. Bovenberg

Yvonne Nygård

Chalmers, Biologi och bioteknik, Industriell bioteknik

ACS Synthetic Biology

2161-5063 (eISSN)

Vol. 5 7 754-64

Ämneskategorier

Annan industriell bioteknik

DOI

10.1021/acssynbio.6b00082