Versatile CRISPR/Cas9 Systems for Genome Editing in Ustilago maydis

Journal of Fungi

Johannes Freitag¹  Sarah-Maria Wege¹  Michael Bölker¹  Fabienne Becker¹  Björn Sandrock¹  Katharina Gejer² 

[1] Department of Biology, Philipps-University Marburg, 35037 Marburg, Germany;Department of Physics, Philipps-University Marburg, 35037 Marburg, Germany;

关键词: Ustilago maydis;    CRISPR/Cas9;    genetic manipulation;    deletion;    point mutation;   

DOI  :  10.3390/jof7020149

来源: DOAJ

【 摘 要 】

The phytopathogenic smut fungus Ustilago maydis is a versatile model organism to study plant pathology, fungal genetics, and molecular cell biology. Here, we report several strategies to manipulate the genome of U. maydis by the CRISPR/Cas9 technology. These include targeted gene deletion via homologous recombination of short double-stranded oligonucleotides, introduction of point mutations, heterologous complementation at the genomic locus, and endogenous N-terminal tagging with the fluorescent protein mCherry. All applications are independent of a permanent selectable marker and only require transient expression of the endonuclease Cas9hf and sgRNA. The techniques presented here are likely to accelerate research in the U. maydis community but can also act as a template for genome editing in other important fungi.

【 授权许可】

Unknown