Trichoderma reesei has been explored intensively in the laboratory and on an industrial scale for its highly potent cellulase secretion machinery since its characterization over 70 years ago. Emergence of new genetic tools over the past decade has strengthened the understanding of mechanism involved in transcription of cellulase genes in fungi and provided a boost to edit them at molecular level. Since several transcriptional factors work synergistically for cellulase expression in fungi; engineering of cellulase secretome for enhanced cellulase titer require combined manipulation of these factors. In the same context, CRISPR/Cas9 has emerged as a powerful, versatile genetic engineering tool for multiplex gene editing in fungi. It is true that considerable efforts with CRISPR technologies have largely developed fungal genetic engineering, but its application in fungi is still challenging and limited. The present review illustrates the precision, strengths and challenges of using CRISPR/Cas9 technology for cellulase engineering in T. reesei, highlighting key strategies that could be employed for strain improvement.

里氏木霉自 70 多年前对其进行表征以来，已在实验室和工业规模上对其高效的纤维素酶分泌机制进行了深入探索。在过去十年中，新的遗传工具的出现加强了对真菌中纤维素酶基因转录机制的理解，并为在分子水平上编辑它们提供了动力。由于几种转录因子协同作用于真菌中的纤维素酶表达；纤维素酶分泌组的工程化以提高纤维素酶滴度需要对这些因素进行综合操作。在同样的背景下，CRISPR/Cas9 已成为一种强大的多功能基因工程工具，用于真菌中的多重基因编辑。的确，在 CRISPR 技术上的巨大努力已经在很大程度上开发了真菌基因工程，但它在真菌中的应用仍然具有挑战性和局限性。本综述说明了使用 CRISPR/Cas9 技术进行纤维素酶工程的精确度、优势和挑战T. reesei，强调了可用于改善应变的关键策略。