ArticleJournal of fungi (Basel, Switzerland)2022
A Mad7 System for Genetic Engineering of Filamentous Fungi.
Article in Journal of fungi (Basel, Switzerland), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 22 citations in OpenAlex.
- Mycoproteins and Beyond: Biotechnological Advances in Filamentous Fungal Biomass.Mini reviews in medicinal chemistry · 2026Article
- Development and validation of PmMAD7 for efficient gene editing in Penaeus monodon.BMC biotechnology · 2025Article
- A CRISPR Cas12a/Cpf1 strategy to facilitate robust multiplex gene editing in Aspergillus Niger.Fungal biology and biotechnology · 2025Article
- Considerations for Domestication of Novel Strains of Filamentous Fungi.ACS synthetic biology · 2025Review
- Expanding the CRISPR Toolbox for Engineering Lycopene Biosynthesis inMicroorganisms · 2024Article
- Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions inJournal of fungi (Basel, Switzerland) · 2024Article
- Cas12a and MAD7, genome editing tools for breeding.Breeding science · 2024Article
- Synthetic Biology Tools for EngineeringJournal of fungi (Basel, Switzerland) · 2024Review
- Utilization of CRISPR-Cas genome editing technology in filamentous fungi: function and advancement potentiality.Frontiers in microbiology · 2024Review
- Strategies for the Development of Industrial Fungal Producing Strains.Journal of fungi (Basel, Switzerland) · 2023Review
- Expanding the genome editing toolbox of Saccharomyces cerevisiae with the endonuclease ErCas12a.FEMS yeast research · 2023Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
Abstract
The introduction of CRISPR technologies has revolutionized strain engineering in filamentous fungi. However, its use in commercial applications has been hampered by concerns over intellectual property (IP) ownership, and there is a need for implementing Cas nucleases that are not limited by complex IP constraints. One promising candidate in this context is the Mad7 enzyme, and we here present a versatile Mad7-CRISPR vector-set that can be efficiently used for the genetic engineering of four different Aspergillus species: Aspergillus nidulans, A. niger, A. oryzae and A. campestris, the latter being a species that has never previously been genetically engineered. We successfully used Mad7 to introduce unspecific as well as specific template-directed mutations including gene disruptions, gene insertions and gene deletions. Moreover, we demonstrate that both single-stranded oligonucleotides and PCR fragments equipped with short and long targeting sequences can be used for efficient marker-free gene editing. Importantly, our CRISPR/Mad7 system was functional in both non-homologous end-joining (NHEJ) proficient and deficient strains. Therefore, the newly implemented CRISPR/Mad7 was efficient to promote gene deletions and integrations using different types of DNA repair in four different Aspergillus species, resulting in the expansion of CRISPR toolboxes in fungal cell factories.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.