Efficient genome editing in rice protoplasts using CRISPR/CAS9 construct

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.). New York : Humana Press, 173-191. (Methods in Molecular Biology, 2238) ISBN 978-1-0716-1070-1

https://doi.org/10.1007/978-1-0716-1068-8_ 11

Chapitre d'ouvrage

Version publiée - Anglais
Accès réservé aux personnels Cirad
Utilisation soumise à autorisation de l'auteur ou du Cirad.
ID606733.pdf
Télécharger (5MB) | Demander une copie

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