The DOI matches its Crossref record; 58 of the 58 references it cites were found in a registry or an index.
A high-throughput gene knockout procedure for Neurospora reveals functions for multiple transcription factors
Abstract
The low rate of homologous recombination exhibited by wild-type strains of filamentous fungi has hindered development of high-throughput gene knockout procedures for this group of organisms. In this study, we describe a method for rapidly creating knockout mutants in which we make use of yeast recombinational cloning, Neurospora mutant strains deficient in nonhomologous end-joining DNA repair, custom-written software tools, and robotics. To illustrate our approach, we have created strains bearing deletions of 103 Neurospora genes encoding transcription factors. Characterization of strains during growth and both asexual and sexual development revealed phenotypes for 43% of the deletion mutants, with more than half of these strains possessing multiple defects. Overall, the methodology, which achieves high-throughput gene disruption at an efficiency >90% in this filamentous fungus, promises to be applicable to other eukaryotic organisms that have a low frequency of homologous recombination.
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A versatile toolkit for high throughput functional genomics with Trichoderma reesei
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Self-citationEfficient Gene Replacements in Toxoplasma gondii Strains Deficient for Nonhomologous End Joining
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Essential Gene Identification and Drug Target Prioritization in Aspergillus fumigatus
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Schumacher, J. (2016) · Molecular Microbiology · doi:10.1111/mmi.13262 · Cited by 157
Metadata
- Authors
- Colot, H. V., Park, G., Turner, G. E., Ringelberg, C., Crew, C. M., Litvinkova, L., Weiss, R. L., Borkovich, K. A., & Dunlap, J. C.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2006-07-05
- Volume
- 103
- Issue
- 27
- Pages
- 10352-10357
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2011485738
- Cited by
- 1,117
- References
- 58
- FWCI
- 12.40
- Type
- journal article
- Language
- en
- Open access
- closed
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- Colot, H. V., Park, G., Turner, G. E., Ringelberg, C., Crew, C. M., Litvinkova, L., Weiss, R. L., Borkovich, K. A., & Dunlap, J. C. (2006). A high-throughput gene knockout procedure for Neurospora reveals functions for multiple transcription factors. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.0601456103
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