CRISPR/Cas9-mediated gene editing in filamentous fungi, focusing on mushrooms and plant-pathogenic fungi

Abstract

CRISPR/Cas gene editing technologies enable precise modifications of the genome and have been extensively applied in genetic studies of animals and plants. Recently, these technologies have been increasingly utilized for the genetic modification of filamentous fungi. However, fungi possess unique cellular characteristics, such as multi-karyosis and heterokaryosis, which pose challenges for efficient gene editing. This review explores these barriers and discusses potential strategies to overcome them. Furthermore, we summarize commonly employed methods for intracellular delivery and expression of Cas9-sgRNA, as well as techniques for selecting transformants. The issue of non-edited nuclei persisting during the regeneration process following transformation is also addressed, along with approaches for their identification and resolution. Finally, we provide an overview of recent advancements in gene editing applications for mushroom-forming filamentous fungi and major plant-pathogenic fungi, focusing key developments in this rapidly evolving field.