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Updated: Jan 25, 2026
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A Fast and Quantitative Method for Post-translational Modification and Variant Enabled Mapping of Peptides to Genomes
Published on: May 22, 2018
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Cpf1 enables fast and efficient genome editing in Aspergilli.
Katherina Garcia Vanegas1, Zofia Dorota Jarczynska1, Tomas Strucko1
1Eukaryotic Molecular Cell Biology, Section for Synthetic Biology, Department of Biotechnology and Biomedicine, Technical University of Denmark, Søltofts Plads, Kongens Lyngby, Denmark.
Fungal Biology and Biotechnology
|May 8, 2019
Summary
We introduce Cpf1, a new CRISPR tool for filamentous fungi. It enables efficient gene editing in Aspergillus species by targeting different DNA sequences than Cas9, expanding CRISPR applications.
Area of Science:
- Molecular Biology
- Genetic Engineering
- Mycology
Background:
- CRISPR technology enhances speed and complexity in fungal genetic engineering.
- Cas9 nuclease is currently the primary CRISPR tool in filamentous fungi, limited by its NGG PAM sequence.
- Alternative RNA-guided nucleases are needed to target a wider range of genomic sites.
Purpose of the Study:
- To introduce and evaluate the Cpf1 nuclease from Lachnospiraceae bacterium (Lb_cpf1) as a novel CRISPR tool for gene editing in filamentous fungi.
- To expand the targeting capabilities of CRISPR technology in fungal species beyond the limitations of Cas9.
Main Methods:
- Codon optimization of Lb_cpf1 for Aspergillus nidulans.
- Development of a vector-based system for Cpf1-mediated CRISPR experiments.
- Application of the system for gene editing in Aspergillus nidulans and Aspergillus niger.
Main Results:
- Codon-optimized Lb_cpf1 functions effectively for CRISPR-based gene editing in filamentous fungi.
- Cpf1-mediated CRISPR experiments were successfully established and demonstrated efficiency at various genomic loci in A. nidulans and A. niger.
- Cpf1 catalyzed oligonucleotide-mediated site-directed mutagenesis and marker-free gene targeting.
Conclusions:
- Cpf1 is presented as a valuable new tool for CRISPR applications in fungi.
- Efficient gene editing using Cpf1 was demonstrated in Aspergillus species.
- The use of Cpf1 expands the range of targetable genomic DNA sequences for CRISPR technologies in fungi.
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