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Int. J. Mol. Sci. 2019, 20(2), 402; https://doi.org/10.3390/ijms20020402

Transgene-Free Genome Editing in Tomato and Potato Plants Using Agrobacterium-Mediated Delivery of a CRISPR/Cas9 Cytidine Base Editor

1
INRA, Agrocampus Ouest, Université Rennes 1, UMR 1349 IGEPP, Domaine de Kéraïber, 29260 Ploudaniel, France
2
INRA PACA, UR 1052, GAFL unit (Génétique et Amélioration des Fruits et Légumes), 84143 Montfavet, France
3
Institut Jean-Pierre Bourgin, INRA, AgroParisTech, CNRS, Université Paris-Saclay, 78000 Versailles, France
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Received: 28 December 2018 / Revised: 14 January 2019 / Accepted: 16 January 2019 / Published: 18 January 2019
(This article belongs to the Special Issue Genome Editing in Plants)
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Abstract

Genome editing tools have rapidly been adopted by plant scientists for gene function discovery and crop improvement. The current technical challenge is to efficiently induce precise and predictable targeted point mutations valuable for crop breeding purposes. Cytidine base editors (CBEs) are CRISPR/Cas9 derived tools recently developed to direct a C-to-T base conversion. Stable genomic integration of CRISPR/Cas9 components through Agrobacterium-mediated transformation is the most widely used approach in dicotyledonous plants. However, elimination of foreign DNA may be difficult to achieve, especially in vegetatively propagated plants. In this study, we targeted the acetolactate synthase (ALS) gene in tomato and potato by a CBE using Agrobacterium-mediated transformation. We successfully and efficiently edited the targeted cytidine bases, leading to chlorsulfuron-resistant plants with precise base edition efficiency up to 71% in tomato. More importantly, we produced 12.9% and 10% edited but transgene-free plants in the first generation in tomato and potato, respectively. Such an approach is expected to decrease deleterious effects due to the random integration of transgene(s) into the host genome. Our successful approach opens up new perspectives for genome engineering by the co-edition of the ALS with other gene(s), leading to transgene-free plants harboring new traits of interest. View Full-Text
Keywords: CRISPR/Cas9; cytidine base editor; Agrobacterium; ALS; transgene-free; potato; tomato CRISPR/Cas9; cytidine base editor; Agrobacterium; ALS; transgene-free; potato; tomato
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Veillet, F.; Perrot, L.; Chauvin, L.; Kermarrec, M.-P.; Guyon-Debast, A.; Chauvin, J.-E.; Nogué, F.; Mazier, M. Transgene-Free Genome Editing in Tomato and Potato Plants Using Agrobacterium-Mediated Delivery of a CRISPR/Cas9 Cytidine Base Editor. Int. J. Mol. Sci. 2019, 20, 402.

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