Genetic barcodes allow traceability of CRISPR/Cas9-derived Aspergillus niger strains without affecting their fitness
| العنوان: | Genetic barcodes allow traceability of CRISPR/Cas9-derived Aspergillus niger strains without affecting their fitness |
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| المؤلفون: | Garrigues, Sandra, Kun, Roland S, de Vries, Ronald P, Sub Molecular Plant Physiology, Molecular Plant Physiology |
| المساهمون: | Westerdijk Fungal Biodiversity Institute, Westerdijk Fungal Biodiversity Institute - Fungal Physiology, Sub Molecular Plant Physiology, Molecular Plant Physiology |
| المصدر: | Current Genetics Current Genetics, 67(4), 673. Springer Verlag Current Genetics, 67(4), 673-684. Springer Verlag GmbH |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | 0106 biological sciences, Antigens, Fungal, Mutant, Computational biology, Biology, Proteomics, 01 natural sciences, 03 medical and health sciences, Genome editing, Gene Expression Regulation, Fungal, 010608 biotechnology, Technical Note, Genetics, Transcription factors, DNA Barcoding, Taxonomic, CRISPR, Cas9, Gene, CRISPR/Cas9, 030304 developmental biology, Barcoding, Risk assessment, Gene Editing, 0303 health sciences, Traceability, General Medicine, Genetically modified organism, Identification (biology), Aspergillus niger, CRISPR-Cas Systems |
| الوصف: | Safe use of genetically modified organisms (GMOs) in biotechnology requires the ability to track the presence of these strains in any environment in which they are applied. For this, introduction of genetic barcodes within the editing site represents a valuable tool for the identification of microbial strains that have undergone genetic modifications. However, it is not known whether these barcodes would have any unexpected effect in the resulting strains or affect the efficiency of the genetic modification. CRISPR/Cas9 has become one of the fastest-growing technologies for genome editing in a range of organisms, including fungi. However, this technology enables the generation of scarless GMOs that are very difficult to distinguish from naturally occurring mutants or other modified organisms. In this study, we address this issue using the industrial workhorse Aspergillus niger as a test case. We applied CRISPR/Cas9 technology to delete the genes encoding the transcriptional regulators XlnR and AraR, involved in the production of plant biomass-degrading enzymes. We generated 20-bp barcoded and non-barcoded ΔxlnR and ΔaraR mutants and analyzed the traceability and fitness of the resulting strains, as well as the efficiency of the genetic modification. Results showed that both barcoded and non-barcoded mutants can be traced by routine PCR reactions when the specific CRISPR/Cas9 modification is known. Additionally, barcodes neither affected the efficiency of the genetic modification nor the growth or protein production of the resulting strains. These results confirm the suitability of genetic barcodes to trace CRISPR-derived GMOs without affecting the performance of the resulting strains. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 0172-8083 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7bc95dfbbecf393b257e67eef9055d9cTest https://doi.org/10.1007/s00294-021-01164-5Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....7bc95dfbbecf393b257e67eef9055d9c |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 01728083 |
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