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Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system

机译：使用CRISPR / Cas9系统高效编辑丝状真菌里氏木霉

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Filamentous fungi have wide applications in biotechnology. The CRISPR/Cas9 system is a powerful genome-editing method that facilitates genetic alterations of genomes in a variety of organisms. However, a genome-editing approach has not been reported in filamentous fungi. Here, we demonstrated the establishment of a CRISPR/Cas9 system in the filamentous fungus Trichoderma reesei by specific codon optimization and in vitro RNA transcription. It was shown that the CRISPR/Cas9 system was controllable and conditional through inducible Cas9 expression. This system generated site-specific mutations in target genes through efficient homologous recombination, even using short homology arms. This system also provided an applicable and promising approach to targeting multiple genes simultaneously. Our results illustrate that the CRISPR/Cas9 system is a powerful genome-manipulating tool for T. reesei and most likely for other filamentous fungal species, which may accelerate studies on functional genomics and strain improvement in these filamentous fungi.
机译：丝状真菌在生物技术中具有广泛的应用。 CRISPR / Cas9系统是一种功能强大的基因组编辑方法，可促进多种生物体中基因组的遗传改变。但是，丝状真菌尚未报道过基因组编辑方法。在这里，我们通过特异性密码子优化和体外RNA转录证明了在丝状真菌里氏木霉中建立了CRISPR / Cas9系统。结果表明，通过诱导Cas9表达，CRISPR / Cas9系统是可控的和有条件的。该系统通过有效的同源重组，即使使用短同源臂，也能在靶基因中产生位点特异性突变。该系统还提供了同时靶向多个基因的适用且有希望的方法。我们的结果表明，CRISPR / Cas9系统是里氏木霉以及其他丝状真菌物种最有力的基因组操纵工具，这可能会加速这些丝状真菌的功能基因组学和菌株改良研究。

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