Genome editing in penicillium chrysogenum using cas9 ribonucleoprotein particles
Book chapter, 2018

Several CRISPR/Cas9 tools have been recently established for precise genome editing in a wide range of filamentous fungi. This genome editing platform offers high flexibility in target selection and the possibility of introducing genetic deletions without the introduction of transgenic sequences. This chapter describes an approach for the transformation of Penicillium chrysogenum protoplasts with preassembled ribonucleoprotein particles (RNPs) consisting of purified Cas9 protein and in vitro transcribed single guide RNA (sgRNA) for the deletion of genome sequences or their replacement with alternative sequences. This method is potentially transferable to all fungal strains where protoplasts can be obtained from.

Ribonucleoprotein particle

HDR

Penicillium chrysogenum

Genome editing

NHEJ

CRISPR/Cas9

RNP

Filamentous fungi

Author

Carsten Pohl

University of Groningen

László Mózsik

University of Groningen

Arnold J.M. Driessen

University of Groningen

R. A. L. Bovenberg

University of Groningen

DSM Biotechnology Centre

Yvonne Nygård

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

University of Groningen

Methods in Molecular Biology

213-232

Subject Categories

Medical Genetics

Bioinformatics and Systems Biology

Genetics

DOI

10.1007/978-1-4939-7795-6_12

More information

Latest update

9/26/2018