ArticleProceedings of the National Academy of Sciences of the United States of America2004
Highly efficient gene replacements in Neurospora strains deficient for nonhomologous end-joining.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 241 papers.
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Who cites it
241 citing papers in PubMed, 613 citations in OpenAlex.
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- Black yeasts are efficient heterologous hosts for the production of a wide range of fungal polyketides.Nature chemical biology · 2026Article
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- A genetic strategy to allow detection of F-actin by phalloidin staining in diverse fungi.mSphere · 2025Article
- Constructing pyrG marker by CRISPR/Cas facilities the highly-efficient precise genome editing on industrial Aspergillus niger strain.Bioprocess and biosystems engineering · 2025Article
- Considerations for Domestication of Novel Strains of Filamentous Fungi.ACS synthetic biology · 2025Review
- Co-transformation of Aspergillus fumigatus: a simple and efficient strategy for gene editing without linking selectable markers.Access microbiology · 2025Article
- Improving the production of micafungin precursor FR901379 inMycology · 2025Article
- A constitutive heterochromatic region shapes genome organization and impacts gene expression in Neurospora crassa.BMC genomics · 2024Article
- Convergent evolution links molybdenum insertase domains with organism-specific sequences.Communications biology · 2024Article
- A Constitutive Heterochromatic Region Shapes Genome Organization and Impacts Gene Expression inbioRxiv : the preprint server for biology · 2024Article
- A user-friendly CRISPR/Cas9 system for mutagenesis of Neurospora crassa.Scientific reports · 2024Article
- NHEJ and HDR can occur simultaneously during gene integration into the genome of Aspergillus niger.Fungal biology and biotechnology · 2024Article
- The First Observation of the Filamentous FungusJournal of fungi (Basel, Switzerland) · 2024Article
- Characterization and regulation of cell cycle-independent noncanonical gene targeting.Nature communications · 2024Article
- Protoplast-mediated transformation of Madurella mycetomatis using hygromycin resistance as a selection marker.PLoS neglected tropical diseases · 2024Article
181 more citing papers are in PubMed but not listed here.
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Gene disruption and overexpression play central roles in the analysis of gene function. Homologous recombination is, in principle, the most efficient method of disrupting, modifying, or replacing a target gene. Although homologous integration of exogenous DNA into the genome occurs readily in Saccharomyces cerevisiae, it is rare in many other organisms. We identified and disrupted Neurospora crassa genes homologous to human KU70 and KU80, which encode proteins that function in nonhomologous end-joining of double-stranded DNA breaks. The resulting mutants, named mus-51 and mus-52, showed higher sensitivity to methyl methanesulfonate, ethyl methanesulfonate, and bleomycin than wild type, but not to UV, 4-nitroquinoline 1-oxide, camptothecin, or hydroxyurea. Vegetative growth, conidiation, and ascospore production in homozygous crosses were normal. The frequency of integration of exogenous DNA into homologous sequences of the genome in the KU disruption strains of N. crassa was compared with that in wild type, mei-3, and mus-11. In mei-3 and mus-11, which are defective in homologous recombination, none or few homologous integration events were observed under any conditions. When mtr target DNA with approximately 2-kb 5' and 3' flanking regions was used for transformation of the KU disruption strains, 100% of transformants exhibited integration at the homologous site, compared to 10 to 30% for a wild-type recipient. Similar results were obtained when the ad-3A gene was targeted for disruption. These results indicate that KU disruption strains are efficient recipients for gene targeting.
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