基于CRISPR/Cas系统的天然产物生物合成基因簇克隆和编辑技术

合成生物学和基因组测序技术的快速发展使挖掘和高效合成天然产物进入了一个全新的时代。由于多数原始菌株生长缓慢、难以培养及遗传改造困难等问题，导致天然产物生物合成基因簇的激活和高效表达受到严重制约。基于此，将原始菌株来源的基因簇转移到操作简便、遗传背景清晰的模式宿主中进行异源表达成为天然产物发现和产量提高的一种有效手段。其中，基因簇的克隆与编辑是实现天然产物异源表达的一个主要限速步骤。CRISPR/Cas技术的应用极大地提高了大型基因簇克隆和编辑的效率，有效促进了微生物来源新药的发现。本文针对基于CRISPR/Cas开发的基因簇克隆和编辑技术进行了系统梳理和全面总结，探讨相关技术在天然产物挖掘和高效合成中的应用及其重要意义。

Cloning and refactoring of natural product biosynthetic gene clusters based on CRISPR/Cas system

Recent rapid developments of synthetic biology and genome sequencing technology opened new possibilities for the discovery and development of microbial natural products. However, a majority of natural products are difficult to access because many microorganisms cannot be cultured, or are difficult to be genetically manipulated. Heterologous expression of biosynthetic gene clusters (BGCs) for natural products in well-characterized hosts provides an alternative way for the discovery of natural products or improvement of their production. However, the success of this strategy hinges upon the cloning and refactoring and/or optimization of large gene clusters. CRISPR/Cas system greatly boosts the development of cloning and refactoring large gene clusters, and promotes the discovery of natural products from microorganisms. In this review, we summarized the recent advances in strategies for gene cluster cloning and refactoring based on CRISPR/Cas system, and discussed the application of related technologies in the discovery and development of natural products and their significance.