Use of transcription activator-like effector for efficient gene modification and transcription in the filamentous fungus Trichoderma reesei.

Abstract

Filamentous fungi have wide applications in the field of biotechnology. The use of transcription activator-like effectors (TALEs) is a powerful genome-engineering tool, which can facilitate genome editing and transcriptional modulation, and has been used for genetic modification in a variety of organisms. However, a transcription activator-like effectors nuclease (TALEN) approach has not been used in filamentous fungi so far. Here, we aimed to establish the shortest TALEN and TALE-transcription factor (TALE-TF) proteins for use in gene modification and transcription in the filamentous fungus Trichoderma reesei. The alternative tandem repeat unit of the TALEs was first established by the ligation of two isocaudamers: XbaI and SpeI, instead of the natural unit. In addition, we adopted T. reesei expression assays to monitor the activities of the TALENs and TALE-TF in vivo in T. reesei. Our results showed that TALEs are a powerful genome-manipulating tool for use in T. reesei and other filamentous fungal species and that their use might facilitate studies on functional genomics and strain improvement in these filamentous fungi.