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Open AccessArticle

Efficient Genome Editing Using CRISPR/Cas9 Technology in Chicory

EA 7394, Institut Charles Viollette (ICV) Agro-food and Biotechnology Research Institute, Université de Lille, INRA, ISA, Univ. Artois, Univ. Littoral Côte d’Opale, Cité Scientifique, 59655 Villeneuve d’Ascq, France
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Int. J. Mol. Sci. 2019, 20(5), 1155; https://doi.org/10.3390/ijms20051155
Received: 30 January 2019 / Revised: 1 March 2019 / Accepted: 2 March 2019 / Published: 6 March 2019
(This article belongs to the Special Issue Genome Editing in Plants)
CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated with protein CAS9) is a genome-editing tool that has been extensively used in the last five years because of its novelty, affordability, and feasibility. This technology has been developed in many plant species for gene function analysis and crop improvement but has never been used in chicory (Cichorium intybus L.). In this study, we successfully applied CRISPR/Cas9-mediated targeted mutagenesis to chicory using Agrobacterium rhizogenes-mediated transformation and protoplast transfection methods. A U6 promoter (CiU6-1p) among eight predicted U6 promoters in chicory was selected to drive sgRNA expression. A binary vector designed to induce targeted mutations in the fifth exon of the chicory phytoene desaturase gene (CiPDS) was then constructed and used to transform chicory. The mutation frequency was 4.5% with the protoplast transient expression system and 31.25% with A. rhizogenes-mediated stable transformation. Biallelic mutations were detected in all the mutant plants. The use of A. rhizogenes-mediated transformation seems preferable as the regeneration of plants is faster and the mutation frequency was shown to be higher. With both transformation methods, foreign DNA was integrated in the plant genome. Hence, selection of vector (transgene)-free segregants is required. Our results showed that genome editing with CRISPR/Cas9 system can be efficiently used with chicory, which should facilitate and accelerate genetic improvement and functional biology. View Full-Text
Keywords: CRISPR/Cas9; multiplex genome editing; Cichorium intybus; phytoene desaturase; Agrobacterium rhizogenes-mediated transformation; protoplast transformation CRISPR/Cas9; multiplex genome editing; Cichorium intybus; phytoene desaturase; Agrobacterium rhizogenes-mediated transformation; protoplast transformation
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MDPI and ACS Style

Bernard, G.; Gagneul, D.; Alves Dos Santos, H.; Etienne, A.; Hilbert, J.-L.; Rambaud, C. Efficient Genome Editing Using CRISPR/Cas9 Technology in Chicory. Int. J. Mol. Sci. 2019, 20, 1155. https://doi.org/10.3390/ijms20051155

AMA Style

Bernard G, Gagneul D, Alves Dos Santos H, Etienne A, Hilbert J-L, Rambaud C. Efficient Genome Editing Using CRISPR/Cas9 Technology in Chicory. International Journal of Molecular Sciences. 2019; 20(5):1155. https://doi.org/10.3390/ijms20051155

Chicago/Turabian Style

Bernard, Guillaume; Gagneul, David; Alves Dos Santos, Harmony; Etienne, Audrey; Hilbert, Jean-Louis; Rambaud, Caroline. 2019. "Efficient Genome Editing Using CRISPR/Cas9 Technology in Chicory" Int. J. Mol. Sci. 20, no. 5: 1155. https://doi.org/10.3390/ijms20051155

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