Wang Duoduo, Jin Shunda, Lu Qianhui, Chen Yupeng
College of Life Sciences, Zhejiang Normal University, Jinhua 321004, China.
Zhejiang Provincial Key Laboratory of Biotechnology on Specialty Economic Plants, Zhejiang Normal University, Jinhua 321004, China.
J Fungi (Basel). 2023 Mar 15;9(3):362. doi: 10.3390/jof9030362.
Fungi represent an important source of bioactive secondary metabolites (SMs), which have wide applications in many fields, including medicine, agriculture, human health, and many other industries. The genes involved in SM biosynthesis are usually clustered adjacent to each other into a region known as a biosynthetic gene cluster (BGC). The recent advent of a diversity of genetic and genomic technologies has facilitated the identification of many cryptic or uncharacterized BGCs and their associated SMs. However, there are still many challenges that hamper the broader exploration of industrially important secondary metabolites. The recent advanced CRISPR/Cas system has revolutionized fungal genetic engineering and enabled the discovery of novel bioactive compounds. In this review, we firstly introduce fungal BGCs and their relationships with associated SMs, followed by a brief summary of the conventional strategies for fungal genetic engineering. Next, we introduce a range of state-of-the-art CRISPR/Cas-based tools that have been developed and review recent applications of these methods in fungi for research on the biosynthesis of SMs. Finally, the challenges and limitations of these CRISPR/Cas-based systems are discussed and directions for future research are proposed in order to expand their applications and improve efficiency for fungal genetic engineering.
真菌是生物活性次生代谢产物（SMs）的重要来源，这些次生代谢产物在许多领域有着广泛应用，包括医学、农业、人类健康以及许多其他行业。参与SM生物合成的基因通常彼此相邻地聚集在一个被称为生物合成基因簇（BGC）的区域。最近多种遗传和基因组技术的出现促进了许多隐秘或未被表征的BGC及其相关SM的鉴定。然而，仍然存在许多挑战阻碍了对具有工业重要性的次生代谢产物的更广泛探索。最近先进的CRISPR/Cas系统彻底改变了真菌基因工程，并使得新型生物活性化合物的发现成为可能。在本综述中，我们首先介绍真菌BGC及其与相关SM的关系，接着简要总结真菌基因工程的传统策略。接下来，我们介绍一系列已开发的基于CRISPR/Cas的先进工具，并回顾这些方法最近在真菌中用于SM生物合成研究的应用。最后，讨论了这些基于CRISPR/Cas的系统的挑战和局限性，并提出了未来研究的方向，以扩大其在真菌基因工程中的应用并提高效率。
