دورية أكاديمية

Genetic barcodes allow traceability of CRISPR/Cas9-derived Aspergillus niger strains without affecting their fitness

التفاصيل البيبلوغرافية
العنوان: Genetic barcodes allow traceability of CRISPR/Cas9-derived Aspergillus niger strains without affecting their fitness
المؤلفون: Garrigues, Sandra, Kun, Roland S., de Vries, Ronald P.
المساهمون: Stichting voor de Technische Wetenschappen
المصدر: Current Genetics ; ISSN 0172-8083 1432-0983
بيانات النشر: Springer Science and Business Media LLC
سنة النشر: 2021
مصطلحات موضوعية: Genetics, General Medicine
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 Δ xlnR and Δ araR 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.
نوع الوثيقة: article in journal/newspaper
اللغة: English
DOI: 10.1007/s00294-021-01164-5
DOI: 10.1007/s00294-021-01164-5.pdf
DOI: 10.1007/s00294-021-01164-5/fulltext.html
الإتاحة: https://doi.org/10.1007/s00294-021-01164-5Test
حقوق: https://creativecommons.org/licenses/by/4.0Test ; https://creativecommons.org/licenses/by/4.0Test
رقم الانضمام: edsbas.170BFB9B
قاعدة البيانات: BASE