ArticleScientific reports2017
Development of a plasmid free CRISPR-Cas9 system for the genetic modification of Mucor circinelloides.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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Who cites it
41 citing papers in PubMed, 90 citations in OpenAlex.
- Advances in molecular tools for elucidating nucleic acid biology in fungal pathogens.microLife · 2026Review
- Three hydrophobic surface binding a proteins link surface hydrophobicity to sporulation, biofilm formation, and host interaction inFrontiers in cellular and infection microbiology · 2026Article
- Calcium starvation leads to strain-specific gene regulation of lipid and carotenoid production in Mucor circinelloides.G3 (Bethesda, Md.) · 2025Article
- Proteomic insights into the physiology and metabolism of oleaginous yeasts and filamentous fungi.Frontiers in microbiology · 2025Review
- Role of the putativeMicrobial cell (Graz, Austria) · 2025Article
- Functional characterization of two survival factor 1 genes inMicrobiology spectrum · 2024Article
- Molecular mechanisms that govern infection and antifungal resistance in Mucorales.Microbiology and molecular biology reviews : MMBR · 2024Review
- Characterization of the Sterol 24-C-Methyltransferase Genes Reveals a Network of Alternative Sterol Biosynthetic Pathways inMicrobiology spectrum · 2023Article
- Industrially Important Fungal Carotenoids: Advancements in Biotechnological Production and Extraction.Journal of fungi (Basel, Switzerland) · 2023Review
- Article
- Application progress of CRISPR/Cas9 genome-editing technology in edible fungi.Frontiers in microbiology · 2023Review
- Mucormycosis in 2023: an update on pathogenesis and management.Frontiers in cellular and infection microbiology · 2023Review
- Manipulation of the Global RegulatorACS chemical biology · 2022Article
- Transcriptome sequencing and global analysis of blue light-responsive genes provide clues for high carotenoid yields in Blakeslea trispora.International microbiology : the official journal of the Spanish Society for Microbiology · 2022Article
- Differential Gene Expression ofJournal of fungi (Basel, Switzerland) · 2022Article
- Development of versatile and efficient genetic tools for the marine-derived fungus Aspergillus terreus RA2905.Current genetics · 2022Article
- Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression.FEMS microbiology letters · 2022Article
- Recent Advances and Future Directions in the Understanding of Mucormycosis.Frontiers in cellular and infection microbiology · 2022Review
- Exploring nano-enabled CRISPR-Cas-powered strategies for efficient diagnostics and treatment of infectious diseases.Journal of nanostructure in chemistry · 2022Review
- Ten decadal advances in fungal biology leading towards human well-being.Fungal diversity · 2022Review
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Mucor circinelloides and other members of Mucorales are filamentous fungi, widely used as model organisms in basic and applied studies. Although genetic manipulation methods have been described for some Mucoral fungi, construction of stable integrative transformants by homologous recombination has remained a great challenge in these organisms. In the present study, a plasmid free CRISPR-Cas9 system was firstly developed for the genetic modification of a Mucoral fungus. The described method offers a rapid but robust tool to obtain mitotically stable mutants of M. circinelloides via targeted integration of the desired DNA. It does not require plasmid construction and its expression in the recipient organism. Instead, it involves the direct introduction of the guide RNA and the Cas9 enzyme and, in case of homology directed repair (HDR), the template DNA into the recipient strain. Efficiency of the method for non-homologous end joining (NHEJ) and HDR was tested by disrupting two different genes, i.e. carB encoding phytoene dehydrogenase and hmgR2 encoding 3-hydroxy-3-methylglutaryl-CoA reductase, of M. circinelloides. Both NHEJ and HDR resulted in stable gene disruption mutants. While NHEJ caused extensive deletions upstream from the protospacer adjacent motif, HDR assured the integration of the deletion cassette at the targeted site.
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