College of Life Sciences, Shanghai Normal University, Shanghai 200234, China.
College of Bioengineering, Henan University of Technology, Zhengzhou 450001, China.
Biomolecules. 2020 May 8;10(5):734. doi: 10.3390/biom10050734.
The genome of encodes a high number of natural product (NP) biosynthetic gene clusters (BGCs). Most of these BGCs are not expressed or are poorly expressed (commonly called silent BGCs) under traditional laboratory experimental conditions. These NP BGCs represent an unexplored rich reservoir of natural compounds, which can be used to discover novel chemical compounds. To activate silent BGCs for NP discovery, two main strategies, including the induction of BGCs expression in native hosts and heterologous expression of BGCs in surrogate hosts, have been adopted, which normally requires genetic manipulation. So far, various genome editing technologies have been developed, which has markedly facilitated the activation of BGCs and NP overproduction in their native hosts, as well as in heterologous hosts. In this review, we summarize the challenges and recent advances in genome editing tools for genetic manipulation with a focus on editing tools based on clustered regularly interspaced short palindrome repeat (CRISPR)/CRISPR-associated protein (Cas) systems. Additionally, we discuss the future research focus, especially the development of endogenous CRISPR/Cas-based genome editing technologies in .
的基因组编码了大量天然产物（NP）生物合成基因簇（BGC）。在传统的实验室实验条件下，这些 BGC 中的大多数没有表达或表达水平较低（通常称为沉默 BGC）。这些 NP BGC 代表了一个未被探索的丰富天然化合物库，可以用于发现新的化合物。为了激活沉默 BGC 以进行 NP 发现，已经采用了两种主要策略，包括在天然宿主中诱导 BGC 表达和在替代宿主中异源表达 BGC，这通常需要遗传操作。到目前为止，已经开发了各种基因组编辑技术，这极大地促进了 BGC 的激活和它们在天然宿主以及异源宿主中的 NP 过量生产。在这篇综述中，我们总结了用于遗传操作的基因组编辑工具的挑战和最新进展，重点介绍了基于成簇规律间隔短回文重复（CRISPR）/CRISPR 相关蛋白（Cas）系统的编辑工具。此外，我们还讨论了未来的研究重点，特别是在 中开发内源性 CRISPR/Cas 基因组编辑技术。
