Advances in CRISPR/Cas9-Based Gene Editing in Filamentous Fungi
As an important class of microorganisms, filamentous fungi have crucial roles in protein secretion, secondary metabolite production and environmental pollution control. However, characteristics such as apical growth, heterokaryon, low homologous recombination (HR) efficiency and the scarcity of gene...
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MDPI AG
2025-05-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/11/5/350 |
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| author | Bin Ma Yimiao Li Tinghui Wang Dongming Li Shuang Jia |
| author_facet | Bin Ma Yimiao Li Tinghui Wang Dongming Li Shuang Jia |
| author_sort | Bin Ma |
| collection | DOAJ |
| description | As an important class of microorganisms, filamentous fungi have crucial roles in protein secretion, secondary metabolite production and environmental pollution control. However, characteristics such as apical growth, heterokaryon, low homologous recombination (HR) efficiency and the scarcity of genetic markers mean that the application of traditional gene editing technology in filamentous fungi faces great challenges. The introduction of the RNA-mediated CRISPR/Cas (clustered regularly interspaced short palindromic repeat/CRlSPR-associated protein) system in filamentous fungi in recent years has revolutionized gene editing in filamentous fungi. In addition, the continuously expressed CRISPR system has significantly improved the editing efficiency, while the optimized sgRNA design and reduced cas9 concentration have effectively reduced the off-target effect, further enhancing the safety and reliability of the technology. In this review, we systematically analyze the molecular mechanism and regulatory factors of CRISPR/Cas9, focus on the optimization of its expression system and the improvement of the transformation efficiency in filamentous fungi, and reveal the core regulatory roles of HR and non-homologous end-joining (NHEJ) pathways in gene editing. Based on the analysis of various filamentous fungi applications, this review reveals the outstanding advantages of CRISPR/Cas9 in the enhancement of protein secretion, addresses the reconstruction of secondary metabolic pathways and pollutant degradation in the past decade, and provides a theoretical basis and practical guidance for the optimization of the technology and engineering applications. |
| format | Article |
| id | doaj-art-016a0889c567419bbdef20c2c6c248b4 |
| institution | Kabale University |
| issn | 2309-608X |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj-art-016a0889c567419bbdef20c2c6c248b42025-08-20T03:47:54ZengMDPI AGJournal of Fungi2309-608X2025-05-0111535010.3390/jof11050350Advances in CRISPR/Cas9-Based Gene Editing in Filamentous FungiBin Ma0Yimiao Li1Tinghui Wang2Dongming Li3Shuang Jia4School of Life Sciences, Inner Mongolia University, Hohhot 010070, ChinaSchool of Life Sciences, Inner Mongolia University, Hohhot 010070, ChinaSchool of Life Sciences, Inner Mongolia University, Hohhot 010070, ChinaSchool of Life Sciences, Inner Mongolia University, Hohhot 010070, ChinaSchool of Life Sciences, Inner Mongolia University, Hohhot 010070, ChinaAs an important class of microorganisms, filamentous fungi have crucial roles in protein secretion, secondary metabolite production and environmental pollution control. However, characteristics such as apical growth, heterokaryon, low homologous recombination (HR) efficiency and the scarcity of genetic markers mean that the application of traditional gene editing technology in filamentous fungi faces great challenges. The introduction of the RNA-mediated CRISPR/Cas (clustered regularly interspaced short palindromic repeat/CRlSPR-associated protein) system in filamentous fungi in recent years has revolutionized gene editing in filamentous fungi. In addition, the continuously expressed CRISPR system has significantly improved the editing efficiency, while the optimized sgRNA design and reduced cas9 concentration have effectively reduced the off-target effect, further enhancing the safety and reliability of the technology. In this review, we systematically analyze the molecular mechanism and regulatory factors of CRISPR/Cas9, focus on the optimization of its expression system and the improvement of the transformation efficiency in filamentous fungi, and reveal the core regulatory roles of HR and non-homologous end-joining (NHEJ) pathways in gene editing. Based on the analysis of various filamentous fungi applications, this review reveals the outstanding advantages of CRISPR/Cas9 in the enhancement of protein secretion, addresses the reconstruction of secondary metabolic pathways and pollutant degradation in the past decade, and provides a theoretical basis and practical guidance for the optimization of the technology and engineering applications.https://www.mdpi.com/2309-608X/11/5/350CRISPR/Cas9filamentous fungigenome editing |
| spellingShingle | Bin Ma Yimiao Li Tinghui Wang Dongming Li Shuang Jia Advances in CRISPR/Cas9-Based Gene Editing in Filamentous Fungi Journal of Fungi CRISPR/Cas9 filamentous fungi genome editing |
| title | Advances in CRISPR/Cas9-Based Gene Editing in Filamentous Fungi |
| title_full | Advances in CRISPR/Cas9-Based Gene Editing in Filamentous Fungi |
| title_fullStr | Advances in CRISPR/Cas9-Based Gene Editing in Filamentous Fungi |
| title_full_unstemmed | Advances in CRISPR/Cas9-Based Gene Editing in Filamentous Fungi |
| title_short | Advances in CRISPR/Cas9-Based Gene Editing in Filamentous Fungi |
| title_sort | advances in crispr cas9 based gene editing in filamentous fungi |
| topic | CRISPR/Cas9 filamentous fungi genome editing |
| url | https://www.mdpi.com/2309-608X/11/5/350 |
| work_keys_str_mv | AT binma advancesincrisprcas9basedgeneeditinginfilamentousfungi AT yimiaoli advancesincrisprcas9basedgeneeditinginfilamentousfungi AT tinghuiwang advancesincrisprcas9basedgeneeditinginfilamentousfungi AT dongmingli advancesincrisprcas9basedgeneeditinginfilamentousfungi AT shuangjia advancesincrisprcas9basedgeneeditinginfilamentousfungi |