Bes Martine, Herbert Leo, Mounier Thibault, Meunier Anne Cecile, Durandet Franz, Guiderdoni Emmanuel, Périn Christophe.
2021. Efficient genome editing in rice protoplasts using CRISPR/CAS9 construct.
In : Rice genome engineering and gene editing: Methods and protocols. Bandyopadhyay Anindya (ed.), Thilmony Roger (ed.)
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Version publiée
- Anglais
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Url - éditeur : https://link.springer.com/protocol/10.1007/978-1-0716-1068-8_11
Résumé : Genome editing technologies, mainly CRISPR/ CAS9, are revolutionizing plant biology and breeding. Since the demonstration of its effectiveness in eukaryotic cells, a very large number of derived technologies has emerged. Demonstrating and comparing the effectiveness of all these new technologies in entire plants is a long, tedious, and labor-intensive process that generally involves the production of transgenic plants and their analysis. Protoplasts, plant cells free of their walls, offer a simple, high -throughput system to test the efficiency of these editing technologies in a few weeks' time span. We have developed a routine protocol using protoplasts to test editing technologies in rice. Our protocol allows to test more than 30 constructs in protoplasts prepared from leaf tissues of 100, 9-11 -day-old seedlings. CRISPR/ CAS9 construct effectiveness can be clearly established within less than a week. We provide here a full protocol, from designing sgRNA to mutation analysis.
Agences de financement hors UE : Agence Nationale de la Recherche
Projets sur financement : (FRA) Ingénierie cellulaire : amélioration et innovation technologiques pour les plantes d'une agriculture
Auteurs et affiliations
- Bes Martine, CIRAD-BIOS-UMR AGAP (FRA)
- Herbert Leo, CIRAD-BIOS-UMR AGAP (FRA)
- Mounier Thibault, CIRAD-BIOS-UMR AGAP (FRA)
- Meunier Anne Cecile, CIRAD-BIOS-UMR AGAP (FRA)
- Durandet Franz, IAGE Company (FRA)
- Guiderdoni Emmanuel, CIRAD-BIOS-UMR AGAP (FRA)
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Périn Christophe, CIRAD-BIOS-UMR AGAP (FRA)
ORCID: 0000-0002-2469-310X
Autres liens de la publication
Source : Cirad-Agritrop (https://agritrop.cirad.fr/606733/)
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