CRISPR/Cas基因编辑及其新兴技术在丝状真菌研究中的系统应用

陈盈盈; 刘扬; 史俊杰; 马俊英; 鞠建华

doi:10.12211/2096-8280.2023-097

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CRISPR/Cas基因编辑及其新兴技术在丝状真菌研究中的系统应用

特约评述 | 更新时间：2025-03-19

- CRISPR/Cas基因编辑及其新兴技术在丝状真菌研究中的系统应用
- CRISPR/Cas systems and their applications in gene editing with filamentous fungi
- 合成生物学 2024年5卷第3期页码：672-693
- 作者机构：
  
  1.中国科学院南海海洋研究所，中国科学院热带海洋生物资源与生态重点实验室，广东省海洋药物重点实验室，广东广州 510301
  2.山东大学药学院，天然产物化学生物学教育部重点实验室，山东省基础科学研究中心（药学），山东省高等学校药物化学生物学重点实验室，山东济南 250012
- 作者简介：
  
  [ "陈盈盈（1986─），女，博士，副研究员。研究方向为真菌次级代谢产物生物合成和代谢调控。E-mail：yychen@scsio.ac.cn" ]
  [ "马俊英（1980─），女，博士，教授，博士生导师。研究方向为结构新颖、活性显著的海洋微生物活性次级代谢产物的生物合成。E-mail：majunying@scsio.ac.cn" ]
  [ "鞠建华（1972─），男，博士，教授，博士生导师。研究方向为微生物活性次级代谢产物的发现、生物合成和抗感染、抗肿瘤创新药物研发。E-mail：jju@sdu.edu.cn" ]
- 基金信息：
  
  国家自然科学基金(22037006;82022067;32300070);广东省自然科学基金(20023A1515011840);广东省重点领域研发计划(2020B111103005);广东省本土创新创业团队项目(2019BT02Y262)
- DOI：10.12211/2096-8280.2023-097
  中图分类号： Q812
- 收稿：2023-12-01，
  
  修回：2024-03-08，
  
  纸质出版：2024-06-30
- 稿件说明：
移动端阅览
陈盈盈，刘扬，史俊杰，马俊英，鞠建华. CRISPR/Cas基因编辑及其新兴技术在丝状真菌研究中的系统应用［J］. 合成生物学， 2024， 5（3）： 672-693

CHEN Yingying， LIU Yang， SHI Junjie， MA Junying， JU Jianhua. CRISPR/Cas systems and their applications in gene editing with filamentous fungi［J］. Synthetic Biology Journal， 2024， 5（3）： 672-693
陈盈盈，刘扬，史俊杰，马俊英，鞠建华. CRISPR/Cas基因编辑及其新兴技术在丝状真菌研究中的系统应用［J］. 合成生物学， 2024， 5（3）： 672-693 DOI： 10.12211/2096-8280.2023-097.

CHEN Yingying， LIU Yang， SHI Junjie， MA Junying， JU Jianhua. CRISPR/Cas systems and their applications in gene editing with filamentous fungi［J］. Synthetic Biology Journal， 2024， 5（3）： 672-693 DOI： 10.12211/2096-8280.2023-097.

摘要

丝状真菌（filamentous fungi）具有独特的形态和细胞构造，与人类健康和工农业生产息息相关，对这类生物资源的开发和利用高度依赖高效的基因编辑平台。然而，由于丝状真菌复杂多样的遗传背景，使用传统的基因编辑技术较难实现大范围的基因编辑，极大地妨碍了丝状真菌的遗传学研究。CRISPR/Cas（clustered regularly interspaced short palindromic repeat/CRISPR-associated protein）技术的出现，打破了这一困境，促进了不同种属和不同来源的丝状真菌的基因编辑，为丝状真菌的基础和应用研究带来了革命性的突破。本文简述了CRISPR/Cas系统的作用机理、分类及基于CRISPR的各种新型技术，归纳总结了丝状真菌中现有的CRISPR/Cas9系统功能组分、多种新兴CRISPR/Cas技术在丝状真菌中的应用现状以及海洋真菌中的CRISPR/Cas技术的应用情况。最后，对CRISPR/Cas系统在丝状真菌中应用进展缓慢、编辑效率低和脱靶效应等问题以及针对这些问题的潜在解决方法进行总结和展望，以期为不同类型的丝状真菌基因编辑平台的构建提供参考。

Abstract

Filamentous fungi

which present distinct morphology and cell structure

play a critical role in human health as well as industrial and agricultural production. However

the unique characteristics of filamentous fungi make them difficult to be manipulated with traditional genetic engineering methods. Thus

the development of an efficient gene editing system is essential for exploring biological resources and understanding metabolic processes in filamentous fungi. The development of the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein (CRISPR/Cas) system promotes more efficient and effective gene editing in different species

and brings a revolutionary breakthrough in fungal fundamental research and applications. In this review

we first briefly introduce the history

working mechanism

and classifications of the CRISPR/Cas mediated gene editing system. Next

we comment the functional components of CRISPR/Cas9 such as selective marker

Cas9 and gRNA and the delivery methods of these components in various filamentous fungi. Furthermore

we systematically discuss the applications of CRISPR related technologies

including CRISPR/Cas12

base-editor

CRISPRa

CRISPRi and CRISPR mediated epigenetic regulation

in the genetic engineering of filamentous fungi

particularly in marine-derived filamentous fungi. Finally

we address challenges with relative low gene editing efficiency and off-targets effects in engineering filamentous fungi

and highlight the potential solutions for developing novel CRISPR/Cas-based gene editing systems. This review can provide guidance for developing an efficient gene editing platform in filamentous fungi and pave the way for further exploration of the secondary metabolites and establishment of robust fungal cell factories.

关键词

Keywords

references

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