CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae

Gene replacements through homologous recombination (HR) have been exten-sively used for functional genomic studies. However, the general efficiency of HR repair can be low in filamentous fungi and the process laborious. Here, we provide a detailed protocol for efficient gene editing by inserting donor DNA into a region of interest following Cas12a ribonucleoprotein (RNP)-mediated DNA double-strand break. We demonstrate this protocol using Magnaporthe or-yzae (synonym of Pyricularia oryzae), a model plant pathogenic fungus that is used to study plant-fungal interactions.For complete details on the use and execution of this protocol, please refer to Huang et al. (2021).

Automated summary

This article presents a detailed protocol for efficient gene editing by inserting donor DNA into a region of interest following Cas12a ribonucleoprotein (RNP)-mediated DNA double-strand break. The protocol is demonstrated using Magnaporthe oryzae, a model plant pathogenic fungus used in the study of plant-fungal interactions.