Induction of targeted deletions in transgenic bread wheat (Triticum aestivum L.) using customized meganuclease
| Autor: | Peter M. Rogowsky, Anne Partier, W. Paul, Caroline Tassy, Pierre Barret, Michel Beckert, D. Youssef, A. Nihou |
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| Přispěvatelé: | Institut National de la Recherche Agronomique (INRA), Génétique Diversité et Ecophysiologie des Céréales (GDEC), Institut National de la Recherche Agronomique (INRA)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), BIOGEMMA, Reproduction et développement des plantes (RDP), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA)-École normale supérieure - Lyon (ENS Lyon), program 'Investments for the Future GENIUS' ANR-11-BTBR-0006-GENIUS, École normale supérieure de Lyon (ENS de Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Transgene [SDV]Life Sciences [q-bio] bread wheat Plant Science Biology Proteomics 01 natural sciences Genome 03 medical and health sciences chemistry.chemical_compound [SDV.BV]Life Sciences [q-bio]/Vegetal Biology biolistics Molecular Biology Gene 2. Zero hunger Genetics Reporter gene gene stacking food and beverages meganuclease genomic DNA 030104 developmental biology chemistry Meganuclease [SDE]Environmental Sciences DNA 010606 plant biology & botany |
| Zdroj: | Plant Molecular Biology Reporter Plant Molecular Biology Reporter, Springer Verlag, 2018, 36 (1), pp.71-81. ⟨10.1007/s11105-017-1062-y⟩ Plant Molecular Biology Reporter, 2018, 36 (1), pp.71-81. ⟨10.1007/s11105-017-1062-y⟩ |
| ISSN: | 0735-9640 |
| Popis: | International audience; Biotechnologies offer breeders good opportunities for breakthrough genetic improvements of bread wheat, one of mankind's main food crops. Since the production of the first transgenic wheat, one of the major concerns has been the removal of selective markers, first because of societal concerns about the antibiotic resistance of some of these genes, and second because removal of a selective marker was the first step toward retransformation using the same selection system. Site-directed nucleases are enzymes that cut genomic DNA in vivo at predefined sites. Among them, meganucleases cut DNA at predefined, long DNA (up to 24 nt) sites, thereby enabling single cuts on large genomes including the bread wheat genome (17 Gbp). In this paper, we describe for the first time the use of a customized meganuclease to cut wheat DNA in vivo. We show that double cuts provoked the deletion of previously inserted DNA cassettes containing the DsRed reporter gene, and that in many cases, the meganuclease target site was correctly reconstituted, offering opportunities for subsequent insertion of stacked transgenes to replace the gene of selection. Moreover, perfect deletions were observed not only in the callus after transient expression of the meganucleases, but also in T0 transgenic wheat after stable retransformation with the meganuclease. Future prospects for the removal of selective markers and transgene stacking are discussed. |
| Databáze: | OpenAIRE |
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