Development of the CRISPR-Cas9 System for the Marine-Derived Fungi Spiromastix sp. SCSIO F190 and Aspergillus sp. SCSIO SX7S7 Article Swipe

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Yingying Chen , Cunlei Cai , Jiafan Yang , Junjie Shi , Yongxiang Song , Dan Hu , Junying Ma , Jianhua Ju ·

YOU? · · 2022 · Open Access · · DOI: https://doi.org/10.3390/jof8070715

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, Spiromastix sp. SCSIO F190 and Aspergillus 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 cas9, 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 rpd3 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.

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Computational Biology

New Mutants

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CRISPR Biology Cas9 Guide RNA Plasmid Gene Transformation (genetics) Genome editing Computational biology Genetics Aspergillus nidulans Mutant

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Type: article
Language: en
Landing Page: https://doi.org/10.3390/jof8070715
PDF: https://www.mdpi.com/2309-608X/8/7/715/pdf?version=1657266434
OA Status: gold
Cited By: 10
References: 50
Related Works: 10
OpenAlex ID: https://openalex.org/W4284962339

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OpenAlex ID: https://openalex.org/W4284962339

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DOI: https://doi.org/10.3390/jof8070715

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Title: Development of the CRISPR-Cas9 System for the Marine-Derived Fungi Spiromastix sp. SCSIO F190 and Aspergillus sp. SCSIO SX7S7

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Type: article

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Language: en

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Publication year: 2022

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Publication date: 2022-07-08

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Authors: Yingying Chen, Cunlei Cai, Jiafan Yang, Junjie Shi, Yongxiang Song, Dan Hu, Junying Ma, Jianhua Ju

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Landing page: https://doi.org/10.3390/jof8070715

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Concepts: CRISPR, Biology, Cas9, Guide RNA, Plasmid, Gene, Transformation (genetics), Genome editing, Computational biology, Genetics, Aspergillus nidulans, Mutant

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Cited by: 10

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abstract_inverted_index.system	43
abstract_inverted_index.target	73
abstract_inverted_index.diverse	13
abstract_inverted_index.firstly	36
abstract_inverted_index.further	161
abstract_inverted_index.genetic	20
abstract_inverted_index.histone	117
abstract_inverted_index.mutated	122
abstract_inverted_index.natural	158
abstract_inverted_index.plasmid	87
abstract_inverted_index.promote	153
abstract_inverted_index.strain.	140
abstract_inverted_index.system,	127
abstract_inverted_index.thereby	128
abstract_inverted_index.(sgRNA),	93
abstract_inverted_index.However,	15
abstract_inverted_index.chemical	41
abstract_inverted_index.circular	86
abstract_inverted_index.emerging	3
abstract_inverted_index.encoding	88
abstract_inverted_index.plasmid.	78
abstract_inverted_index.produced	136
abstract_inverted_index.products	159
abstract_inverted_index.resulted	98
abstract_inverted_index.strains.	113
abstract_inverted_index.strategy	69
abstract_inverted_index.studies.	163
abstract_inverted_index.targeted	104
abstract_inverted_index.addition,	115
abstract_inverted_index.compounds	27
abstract_inverted_index.developed	61
abstract_inverted_index.discovery	24, 155
abstract_inverted_index.effective	39
abstract_inverted_index.efficient	19
abstract_inverted_index.frequency	102
abstract_inverted_index.mutations	108
abstract_inverted_index.producers	6
abstract_inverted_index.secondary	10, 131
abstract_inverted_index.together,	142
abstract_inverted_index.versatile	65, 144
abstract_inverted_index.wild-type	139
abstract_inverted_index.activating	129
abstract_inverted_index.attractive	5
abstract_inverted_index.biological	162
abstract_inverted_index.disruption	68, 147
abstract_inverted_index.selectable	96
abstract_inverted_index.Aspergillus	55
abstract_inverted_index.CRISPR-Cas9	126
abstract_inverted_index.Spiromastix	50
abstract_inverted_index.deacetylase	118
abstract_inverted_index.elucidation	30
abstract_inverted_index.established	37, 125
abstract_inverted_index.insertional	106
abstract_inverted_index.mechanisms.	33
abstract_inverted_index.metabolites	11, 132
abstract_inverted_index.protoplasts	45
abstract_inverted_index.PEG-mediated	40
abstract_inverted_index.biosynthesis	32
abstract_inverted_index.established,	150
abstract_inverted_index.single-guide	91
abstract_inverted_index.structurally	8
abstract_inverted_index.transforming	71
abstract_inverted_index.Marine-derived	0
abstract_inverted_index.bioactivities.	14
abstract_inverted_index.marine-derived	48, 111
abstract_inverted_index.transformation	42, 83
abstract_inverted_index.CRISPR-Cas9-based	66, 145
cited_by_percentile_year.max	98
cited_by_percentile_year.min	94
corresponding_author_ids	https://openalex.org/A5028227890, https://openalex.org/A5090344676
countries_distinct_count	1
institutions_distinct_count	8
corresponding_institution_ids	https://openalex.org/I19820366, https://openalex.org/I4210101167, https://openalex.org/I4210103685, https://openalex.org/I4210106122
sustainable_development_goals[0].id	https://metadata.un.org/sdg/14
sustainable_development_goals[0].score	0.8799999952316284
sustainable_development_goals[0].display_name	Life below water
citation_normalized_percentile.value	0.73298616
citation_normalized_percentile.is_in_top_1_percent	False
citation_normalized_percentile.is_in_top_10_percent	False