Genome Editing Technology and Its Application Potentials in the Industrial Filamentous Fungus Aspergillus oryzae

Maruyama, Jun‐ichi

doi:10.3390/jof7080638

Cited by 19 publications

(8 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One reason could be that the vectors containing CRISPR-related elements were not delivered efficiently. There are few reports on autonomous replicators, except for AMA1 [ 41 , 42 ], which is only found in Aspergillus. Hence, CRISPR-related components will duplicate along with genome duplication.…”

Section: Discussionmentioning

confidence: 99%

Establishment of CRISPR/Cas9 Genome-Editing System Based on Dual sgRNAs in Flammulina filiformis

Liu

Dong

Liao

et al. 2022

JoF

View full text Add to dashboard Cite

Flammulina filiformis, previously known as Asian Flammulina velutipes, is one of the most commercially important edible fungi, with nutritional value and medicinal properties worldwide. However, precision genome editing using CRISPR/Cas9, which is a revolutionary technology and provides a powerful tool for molecular breeding, has not been established in F. filiformis. Here, plasmids harboring expression cassettes of Basidiomycete codon-optimized Cas9 and dual sgRNAs targeting pyrG under the control of the gpd promoter and FfU6 promoter, respectively, were delivered into protoplasts of F. filiformis Dan3 strain through PEG-mediated transformation. The results showed that an efficient native U6 promoter of F. filiformis was identified, and ultimately several pyrG mutants exhibiting 5-fluorooric acid (5-FOA) resistance were obtained. Additionally, diagnostic PCR followed by Sanger sequencing revealed that fragment deletion between the two sgRNA target sites or small insertions and deletions (indels) were introduced in these pyrG mutants through the nonhomologous end joining (NHEJ) pathway, resulting in heritable changes in genomic information. Taken together, this is the first report in which a successful CRISPR/Cas9 genome-editing system based on dual sgRNAs was established in F. filiformis, which broadens the application of this advanced tool in Basidiomycetes.

show abstract

Section: Discussionmentioning

confidence: 99%

Establishment of CRISPR/Cas9 Genome-Editing System Based on Dual sgRNAs in Flammulina filiformis

Liu

Dong

Liao

et al. 2022

JoF

View full text Add to dashboard Cite

show abstract

“…Conversely, HR relies on undamaged homologous regions as templates to mend DSBs with precise insertions or deletions, thereby achieving gene knockouts (Figure 2a). TALENs and CRISPR/Cas9 are pivotal gene editing enzymes in A. oryzae genetic manipulation research [69]. TALENs consist of transcription activator-like effectors (TALEs) and the FokI nuclease.…”

Section: Genome Editing Techniquesmentioning

confidence: 99%

Synthetic Biology Tools for Engineering Aspergillus oryzae

Yang,

Song,

Liu

et al. 2024

JoF

View full text Add to dashboard Cite

For more than a thousand years, Aspergillus oryzae has been used in traditional culinary industries, including for food fermentation, brewing, and flavoring. In recent years, A. oryzae has been extensively used in deciphering the pathways of natural product synthesis and value-added compound bioproduction. Moreover, it is increasingly being used in modern biotechnology industries, such as for the production of enzymes and recombinant proteins. The investigation of A. oryzae has been significantly accelerated through the successive application of a diverse array of synthetic biology techniques and methodologies. In this review, the advancements in biological tools for the synthesis of A. oryzae, including DNA assembly technologies, gene expression regulatory elements, and genome editing systems, are discussed. Additionally, the challenges associated with the heterologous expression of A. oryzae are addressed.

show abstract

“…A. oryzae has been known to have a strong ability to secrete large amounts of hydrolytic enzymes, and this property has been widely exploited in the production of recombinant proteins and secondary metabolites [ 86 ]. In recent years, the CRISPR/Cas9 system, a versatile genomic editing technology, has been rapidly developed in A. oryzae to better adapt to its application in industrial production [ 87 ]. Katayama et al [ 48 ] were the first to establish CRISPR/Cas9-based genome editing in A. oryzae successfully.…”

Section: Development and Application Of Crispr/cas9-based Genome Edit...mentioning

confidence: 99%

CRISPR/Cas9-Based Genome Editing and Its Application in Aspergillus Species

Jin

Wang

et al. 2022

JoF

View full text Add to dashboard Cite

Aspergillus, a genus of filamentous fungi, is extensively distributed in nature and plays crucial roles in the decomposition of organic materials as an important environmental microorganism as well as in the traditional fermentation and food processing industries. Furthermore, due to their strong potential to secrete a large variety of hydrolytic enzymes and other natural products by manipulating gene expression and/or introducing new biosynthetic pathways, several Aspergillus species have been widely exploited as microbial cell factories. In recent years, with the development of next-generation genome sequencing technology and genetic engineering methods, the production and utilization of various homo-/heterologous-proteins and natural products in Aspergillus species have been well studied. As a newly developed genome editing technology, the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system has been used to edit and modify genes in Aspergilli. So far, the CRISPR/Cas9-based approach has been widely employed to improve the efficiency of gene modification in the strain type Aspergillus nidulans and other industrially important and pathogenic Aspergillus species, including Aspergillus oryzae, Aspergillus niger, and Aspergillus fumigatus. This review highlights the current development of CRISPR/Cas9-based genome editing technology and its application in basic research and the production of recombination proteins and natural products in the Aspergillus species.

show abstract

Genome Editing Technology and Its Application Potentials in the Industrial Filamentous Fungus Aspergillus oryzae

Cited by 19 publications

References 40 publications

Establishment of CRISPR/Cas9 Genome-Editing System Based on Dual sgRNAs in Flammulina filiformis

Establishment of CRISPR/Cas9 Genome-Editing System Based on Dual sgRNAs in Flammulina filiformis

Synthetic Biology Tools for Engineering Aspergillus oryzae

CRISPR/Cas9-Based Genome Editing and Its Application in Aspergillus Species

Contact Info

Product

Resources

About