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Expanding the CRISPR Toolbox in P. patens Using SpCas9-NG Variant and Application for Gene and Base Editing in Solanaceae Crops

Authors :
Veillet, Florian
Perrot, Laura
Guyon-Debast, Anouchka
Kermarrec, Marie-Paule
Chauvin, Laura
Chauvin, Jean-Eric
Gallois, Jean-Luc
Mazier, Marianne
Nogué, Fabien
Institut de Génétique, Environnement et Protection des Plantes (IGEPP)
Université de Rennes 1 (UR1)
Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-AGROCAMPUS OUEST
Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Génétique et Amélioration des Fruits et Légumes (GAFL)
Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Institut Jean-Pierre Bourgin (IJPB)
AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
ANR-10-LABX-0040-SPS, Agence Nationale de la Recherche
ANR-11-BTBR-0001,GENIUS,Ingénierie cellulaire : amélioration et innovation technologiques pour les plantes d'une agriculture(2011)
ANR-17-EURE-0007,SPS-GSR,Ecole Universitaire de Recherche de Sciences des Plantes de Paris-Saclay(2017)
Université de Rennes (UR)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-INSTITUT AGRO Agrocampus Ouest
Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)
Source :
International Journal of Molecular Sciences, International Journal of Molecular Sciences, MDPI, 2020, 21 (3), pp.1024. ⟨10.3390/ijms21031024⟩, International Journal of Molecular Sciences, Vol 21, Iss 3, p 1024 (2020), International Journal of Molecular Sciences, 2020, 21 (3), pp.1024. ⟨10.3390/ijms21031024⟩
Publication Year :
2020
Publisher :
HAL CCSD, 2020.

Abstract

Genome editing has become a major tool for both functional studies and plant breeding in several species. Besides generating knockouts through the classical CRISPR-Cas9 system, recent development of CRISPR base editing holds great and exciting opportunities for the production of gain-of-function mutants. The PAM requirement is a strong limitation for CRISPR technologies such as base editing, because the base substitution mainly occurs in a small edition window. As precise single amino-acid substitution can be responsible for functions associated to some domains or agronomic traits, development of Cas9 variants with relaxed PAM recognition is of upmost importance for gene function analysis and plant breeding. Recently, the SpCas9-NG variant that recognizes the NGN PAM has been successfully tested in plants, mainly in monocotyledon species. In this work, we studied the efficiency of SpCas9-NG in the model moss Physcomitrella patens and two Solanaceae crops (Solanum lycopersicum and Solanum tuberosum) for both classical CRISPR-generated gene knock-out and cytosine base editing. We showed that the SpCas9-NG greatly expands the scope of genome editing by allowing the targeting of non-canonical NGT and NGA PAMs. The CRISPR toolbox developed in our study opens up new gene function analysis and plant breeding perspectives for model and crop plants.

Details

Language :
English
ISSN :
16616596 and 14220067
Database :
OpenAIRE
Journal :
International Journal of Molecular Sciences, International Journal of Molecular Sciences, MDPI, 2020, 21 (3), pp.1024. ⟨10.3390/ijms21031024⟩, International Journal of Molecular Sciences, Vol 21, Iss 3, p 1024 (2020), International Journal of Molecular Sciences, 2020, 21 (3), pp.1024. ⟨10.3390/ijms21031024⟩
Accession number :
edsair.dedup.wf.001..2a217a0387e04b45ed704d0806311919