Development of the CRISPR-Cas9 System for the Marine-Derived Fungi <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7
Marine-derived fungi are emerging as attractive producers of structurally novel secondary metabolites with diverse bioactivities. However, the lack of efficient genetic tools limits the discovery of novel compounds and the elucidation of biosynthesis mechanisms. Here, we firstly established an effec...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-07-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/8/7/715 |
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| author | Yingying Chen Cunlei Cai Jiafan Yang Junjie Shi Yongxiang Song Dan Hu Junying Ma Jianhua Ju |
| author_facet | Yingying Chen Cunlei Cai Jiafan Yang Junjie Shi Yongxiang Song Dan Hu Junying Ma Jianhua Ju |
| author_sort | Yingying Chen |
| collection | DOAJ |
| description | Marine-derived fungi are emerging as attractive producers of structurally novel secondary metabolites with diverse bioactivities. However, the lack of efficient genetic tools limits the discovery of novel compounds and the elucidation of biosynthesis mechanisms. Here, we firstly established an effective PEG-mediated chemical transformation system for protoplasts in two marine-derived fungi, <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7. Next, we developed a simple and versatile CRISPR-Cas9-based gene disruption strategy by transforming a target fungus with a single plasmid. We found that the transformation with a circular plasmid encoding <i>cas9</i>, a single-guide RNA (sgRNA), and a selectable marker resulted in a high frequency of targeted and insertional gene mutations in both marine-derived fungal strains. In addition, the histone deacetylase gene <i>rpd3</i> was mutated using the established CRISPR-Cas9 system, thereby activating novel secondary metabolites that were not produced in the wild-type strain. Taken together, a versatile CRISPR-Cas9-based gene disruption method was established, which will promote the discovery of novel natural products and further biological studies. |
| first_indexed | 2024-03-09T13:34:06Z |
| format | Article |
| id | doaj.art-2f1dc8492f064ca59d173d98b0d02ddd |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-09T13:34:06Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-2f1dc8492f064ca59d173d98b0d02ddd2023-11-30T21:14:20ZengMDPI AGJournal of Fungi2309-608X2022-07-018771510.3390/jof8070715Development of the CRISPR-Cas9 System for the Marine-Derived Fungi <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7Yingying Chen0Cunlei Cai1Jiafan Yang2Junjie Shi3Yongxiang Song4Dan Hu5Junying Ma6Jianhua Ju7CAS Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, ChinaGuangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, Institute of Traditional Chinese Medicine and Natural Products, College of Pharmacy, Jinan University, Guangzhou 510632, ChinaCAS Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, ChinaMarine-derived fungi are emerging as attractive producers of structurally novel secondary metabolites with diverse bioactivities. However, the lack of efficient genetic tools limits the discovery of novel compounds and the elucidation of biosynthesis mechanisms. Here, we firstly established an effective PEG-mediated chemical transformation system for protoplasts in two marine-derived fungi, <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7. Next, we developed a simple and versatile CRISPR-Cas9-based gene disruption strategy by transforming a target fungus with a single plasmid. We found that the transformation with a circular plasmid encoding <i>cas9</i>, a single-guide RNA (sgRNA), and a selectable marker resulted in a high frequency of targeted and insertional gene mutations in both marine-derived fungal strains. In addition, the histone deacetylase gene <i>rpd3</i> was mutated using the established CRISPR-Cas9 system, thereby activating novel secondary metabolites that were not produced in the wild-type strain. Taken together, a versatile CRISPR-Cas9-based gene disruption method was established, which will promote the discovery of novel natural products and further biological studies.https://www.mdpi.com/2309-608X/8/7/715marine-derived fungiCRISPR-Cas9<i>Spiromastix</i> sp. SCSIO F190<i>Aspergillus</i> sp. SCSIO SX7S7secondary metaboliteshistone deacetylase |
| spellingShingle | Yingying Chen Cunlei Cai Jiafan Yang Junjie Shi Yongxiang Song Dan Hu Junying Ma Jianhua Ju Development of the CRISPR-Cas9 System for the Marine-Derived Fungi <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7 Journal of Fungi marine-derived fungi CRISPR-Cas9 <i>Spiromastix</i> sp. SCSIO F190 <i>Aspergillus</i> sp. SCSIO SX7S7 secondary metabolites histone deacetylase |
| title | Development of the CRISPR-Cas9 System for the Marine-Derived Fungi <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7 |
| title_full | Development of the CRISPR-Cas9 System for the Marine-Derived Fungi <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7 |
| title_fullStr | Development of the CRISPR-Cas9 System for the Marine-Derived Fungi <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7 |
| title_full_unstemmed | Development of the CRISPR-Cas9 System for the Marine-Derived Fungi <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7 |
| title_short | Development of the CRISPR-Cas9 System for the Marine-Derived Fungi <i>Spiromastix</i> sp. SCSIO F190 and <i>Aspergillus</i> sp. SCSIO SX7S7 |
| title_sort | development of the crispr cas9 system for the marine derived fungi i spiromastix i sp scsio f190 and i aspergillus i sp scsio sx7s7 |
| topic | marine-derived fungi CRISPR-Cas9 <i>Spiromastix</i> sp. SCSIO F190 <i>Aspergillus</i> sp. SCSIO SX7S7 secondary metabolites histone deacetylase |
| url | https://www.mdpi.com/2309-608X/8/7/715 |
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