CRISPR/Cas9编辑系统在微生物天然产物研究中的应用
Applications of the CRISPR/Cas9 editing system in the study of microbial natural products
- 合成生物学 2024年5卷第3期 页码：658-671
- 作者机构：
1.香港科技大学理学院化学系，清水湾校区，香港 999077
2.深圳湾实验室，化学生物学研究所，广东 深圳 518132
- 作者简介：
[ "惠真 （1989—）， 男， 博士研究生。研究方向为微生物天然产物基因挖掘和生物合成。E-mail：zhuiaa@connect.ust.hk" ]
[ "唐啸宇（1984—），男，博士，研究员，博士生导师。研究方向为微生物天然产物化学生物学。E-mail：xtang@szbl.ac.cn" ]
- 基金信息：国家自然科学基金面上项目(82173719)
DOI：10.12211/2096-8280.2023-110
中图分类号： Q812收稿：2023-12-26，
修回：2024-03-17，
纸质出版：2024-06-30
- 稿件说明：
移动端阅览
惠真， 唐啸宇. CRISPR/Cas9编辑系统在微生物天然产物研究中的应用［J］. 合成生物学， 2024， 5（3）： 658-671
HUI Zhen， TANG Xiaoyu. Applications of the CRISPR/Cas9 editing system in the study of microbial natural products［J］. Synthetic Biology Journal， 2024， 5（3）： 658-671
惠真， 唐啸宇. CRISPR/Cas9编辑系统在微生物天然产物研究中的应用［J］. 合成生物学， 2024， 5（3）： 658-671 DOI： 10.12211/2096-8280.2023-110.
HUI Zhen， TANG Xiaoyu. Applications of the CRISPR/Cas9 editing system in the study of microbial natural products［J］. Synthetic Biology Journal， 2024， 5（3）： 658-671 DOI： 10.12211/2096-8280.2023-110.
摘要
微生物作为天然产物的巨大宝库，一直以来都是研究人员挖掘和开发新的活性化合物的重要来源。目前，利用基因编辑工具发现、生物合成和代谢调控天然产物的研究方法受到该领域研究者的广泛关注。CRISPR/Cas9遗传编辑系统以其独特的灵活靶向优势克服了其他遗传编辑方法常见的对序列同源或位点限制，简化了实验步骤，提高了实验效率，促进了天然产物研究领域的发展。本文主要介绍CRISPR/Cas9系统在微生物天然产物发现、生物合成和工程改造方面的应用，分别从CRISPR/Cas9系统的发展、天然产物生物合成基因簇的克隆和遗传编辑、天然产物结构衍生化和代谢调节、沉默天然产物基因簇的激活这几个方面阐述CRISPR/Cas9系统在微生物天然产物研究领域的优势。最后，针对CRISPR/Cas9系统无法克服的重组效率和宿主适应性问题提供了可行的解决思路。相信随着合成生物学和信息技术的发展，越来越多的与CRISPR/Cas9系统相关的遗传操作工具和方法会被开发，将不断推动天然产物领域的发展进步。
Abstract
Microorganisms have consistently been a crucial source for researchers to explore and develop new natural products. Currently
research methods involving gene editing tools for the discovery
biosynthesis
and metabolic engineering of natural products have garnered broad attention in this field. However
traditional methods for gene editing usually rely on the recombination ability of the host or introduced proteins. It’s difficult to establish a general platform for all bacteria mainly because of their complicated genetic background. This genetic diversity often causes laborious experimental operations with low efficiency. The CRISPR/Cas9 gene editing system
with its unique and flexible targeting advantages
overcomes common limitations such as sequence homology or site constraint in other gene editing methods and thus is more likely to function in diverse bacteria species. This simplifies experimental procedures
enhances work efficiency
and promotes the development of natural product research. This article introduces the applications of the CRISPR/Cas9 system for the discovery
biosynthesis
and metabolic engineering of natural products in microorganisms. It covers the development of the CRISPR/Cas9 system
cloning and genetic editing of natural product biosynthetic gene clusters
structural derivatization and metabolic engineering of natural products
and the activation of silenced natural product biosynthetic gene clusters. These aspects highlight the advantages of the CRISPR/Cas9 system in the research of natural products with microorganisms. Finally
solutions are proposed for addressing challenges that the CRISPR/Cas9 system currently faces in overcoming low recombination efficiency and host adaptability issues. Especially the CRISPR/Cas12a system which has broadened applications of the CRISRP/Cas9 system by preferring different PAM sites. In addition to functions that CRISPR/Cas9 system has realized
its potent multiple targeting ability further enhances the efficiency of target editing. It is believed that with the development of synthetic biology and information technology
an increasing number of genetic manipulation tools and methods related to the CRISPR/Cas9 system will be developed
continually driving progress in the research of natural products.
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关键词
Keywords
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