工业丝状真菌基因组编辑技术研究进展

刘倩; 李金根; 张晨阳; 李芳雅; 田朝光

doi:10.12211/2096-8280.2020-073

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工业丝状真菌基因组编辑技术研究进展

特约评述 | 更新时间：2025-03-20

- 工业丝状真菌基因组编辑技术研究进展
- Research progress of genome editing technologies for industrial filamentous fungi
- 合成生物学 2021年2卷第2期页码：256-273
- 作者机构：
  
  1.中国科学院天津工业生物技术研究所，系统微生物工程重点实验室，天津 300308
  2.国家合成生物技术创新中心，天津 300308
  3.中国科学院大学，北京 100049
- 作者简介：
  
  [ "刘倩（1983─），女，博士，副研究员，研究方向为工业真菌遗传改造。E-mail：liu_q1@tib.cas.cn" ]
  [ "田朝光（1973─），男，博士，研究员，研究方向为真菌合成生物学。E-mail：tian_cg@tib.cas.cn" ]
- 基金信息：
  
  国家重点研发计划“合成生物学专项”(2018FYA0900500);天津市合成生物技术创新能力提升行动(TSBICIP-KJGG-006);国家自然科学基金项目(31972878;31771386;31972879);中国科学院青年创新促进会(2019180;2020183)
- DOI：10.12211/2096-8280.2020-073
  中图分类号： Q812
- 收稿：2020-07-09，
  
  修回：2021-03-15，
  
  纸质出版：2021-04-30
- 稿件说明：
移动端阅览
刘倩, 李金根, 张晨阳, 李芳雅, 田朝光. 工业丝状真菌基因组编辑技术研究进展[J]. 合成生物学, 2021, 2(2): 256-273

LIU Qian, LI Jingen, ZHANG Chenyang, LI Fangya, TIAN Chaoguang. Research progress of genome editing technologies for industrial filamentous fungi[J]. Synthetic Biology Journal, 2021, 2(2): 256-273
刘倩, 李金根, 张晨阳, 李芳雅, 田朝光. 工业丝状真菌基因组编辑技术研究进展[J]. 合成生物学, 2021, 2(2): 256-273 DOI： 10.12211/2096-8280.2020-073.

LIU Qian, LI Jingen, ZHANG Chenyang, LI Fangya, TIAN Chaoguang. Research progress of genome editing technologies for industrial filamentous fungi[J]. Synthetic Biology Journal, 2021, 2(2): 256-273 DOI： 10.12211/2096-8280.2020-073.

摘要

丝状真菌（filamentous fungi）是广泛存在于自然界中的一类多细胞真核微生物，是酶制剂、有机酸、抗生素的核心生产体系，在生物技术中发挥着非常重要的作用。由于丝状真菌的生长发育较为复杂，其遗传体系和基因组编辑技术发展相对较慢，妨碍了丝状真菌基础研究和开发利用的快速发展。近年来，核酸酶介导的基因组编辑技术已经发展成为一种功能强大的基因组编辑工具，在生物技术领域的应用得到广泛关注，其中RNA介导的CRISPR系统（clustered regularly interspaced short palindromic repeats）已经成为新一代基因组编辑技术体系。文章将对目前使用最为广泛的3种基因编辑系统，包括CRISPR-Cas技术的发展历程进行概述，对CRISPR-Cas技术在丝状真菌基因组编辑中的研发进行系统介绍，包括在工业丝状真菌中的研发应用，例如鉴定次级代谢产物的关键基因并提高次级代谢物的生产能力、将外源基因定点整合到基因组上从而提高异源蛋白的表达、遗传重构蛋白分泌途径提高工业酶的产量等，最后对CRISPR-Cas基因组编辑技术及其衍生系统在丝状真菌的真菌基因功能研究、代谢途径重构、精确表达调控、蛋白定向进化以及高性能底盘构建等方面进行了展望。

Abstract

Filamentous fungi

which are a large and diverse group of multicellular eukaryotic microorganisms existing in nature universally

have played pivotal roles in biotechnology as the prominent producers of enzymes

organic acids and antibiotics. Filamentous fungi are also important decomposers that contribute to the biological carbon cycle of plant biomass. Genetic engineering is a powerful approach for researchers not only to elucidate the gene function in filamentous fungi

but also to improve their production levels and minimize unwanted by-product formation. However

the efficiency of homologous integration in filamentous fungi is very low using classical genetic approaches. Due to the complicated growth and lifestyle of filamentous fungi

the development of genetic tools and gene editing is relatively slow

which hinders the basic research and biotechnological development of filamentous fungi. In recent years

genome editing technologies based on programmable nucleases have been developed as the powerful gene engineering tools in a wide variety of organisms. The most rapidly developed and wildly used technology is the class of RNA-guided Cas9 nuclease known as the adaptive immune system CRISPR (clustered regularly interspaced short palindromic repeats). The Cas9-sgRNA complex binds to the corresponding target site of the protospacer in a genome

and the specifically induces double-strand breaks. These breaks can be used as a basis for site-specific mutagenesis mediated by non-homologous end-joining or for the introduction of precise mutation or integration via homology-directed repair. CRISPR-Cas systems have recently enabled a wide range of applications for genome editing in many organisms. Remarkably

and in just the past few years

the CRISPR-Cas9 system has emerged as a more efficient strategy for gene editing in filamentous fungi. In this review

we describe the research progress of three most widely used genome editing systems

including the development of CRISPR-Cas technology and its applications in filamentous fungi

especially recent advances in industrial filamentous fungi. Finally

we give the perspectives for the CRISPR-Cas technology and its derivative systems for genomic editing in filamentous fungi.

关键词

Keywords

references

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