Gene editing in M. oryzae was performed using purified Cas9 pre-complexed to RNA guides to form ribonucleoproteins (RNPs).

Gene editing in M. oryzae was performed using purified Cas9 pre-complexed to RNA guides to form ribonucleoproteins (RNPs). When used in combination with oligonucleotide or PCR-generated donor DNAs, the generation of strains with specific base pair edits, in-locus gene replacements, or multiple gene edits, is very rapid and straightforward. Additionally, we report a novel counterselection strategy which allows creation of precisely edited fungal strains that contain no foreign DNA and are completely isogenic to the wild type.