Targeted Gene Disruption Via CRISPR/Cas9 Ribonucleoprotein Complexes in Fusarium oxysporum

Characterization of a gene of interest frequently relies on generation of a mutant as a critical component. Transformation to disrupt a gene has been previously accomplished by several methods in Fusarium oxysporum. Here we provide a detailed method to generate a gene mutation mediated by a CRISPR/Cas9 ribonucleoprotein (RNP) complex. The Cas9 RNP cleaves the DNA at the target site, and during DNA repair integration of a dominant selectable marker is incorporated via homologous recombination generating the desired gene disruption.