Tailor‐made TALEN system for highly efficient targeted gene replacement in the rice blast fungus

Abstract Genetic manipulation is key to unraveling gene functions and creating genetically modified strains of microbial organisms. Recently, engineered nucleases that can generate DNA double‐strand breaks (DSBs) at a specific site in the desired locus within genome are utilized in a rapidly developing genome editing technology via DSBs repair. However, the use of engineered nucleases in filamentous fungi has not been validated. In this study, we demonstrated that tailor‐made transcriptional activator‐like effector nucleases (TALENs) system, Platinum–Fungal TALENs (PtFg TALENs), could improve the efficiency of homologous recombination‐mediated targeted gene replacement by up to 100% in the rice blast fungus Pyricularia oryzae . This high‐efficiency PtFg TALEN has great potential for basic and applied biological applications in filamentous fungi. Biotechnol. Bioeng. 2015;112: 1335–1342. © 2015 Wiley Periodicals, Inc.