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. 2019 Feb 6;10:40. doi: 10.3389/fpls.2019.00040

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Copyright © 2019 Charrier, Vergne, Dousset, Richer, Petiteau and Chevreau.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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CRISPR-Cas expression construct. The Cas9 gene from Streptococcus pyogenes is driven by PcUbi4-2 promoter (P) from parsley (Petroselinum crispum) and transcription is terminated by the Pea3a terminator (T) from pea (Pisum sativum). gRNA1 and 2 are, respectively, driven by MdU3 and MdU6 promoters from Malus domestica and transcription is terminated by a polyT terminator. Transformants are selected with a nptII gene controlled by nos promoter and terminator from Agrobacterium tumefaciens. AttB1 and 2: sites resulting from the Gateway^® LR recombination. LB and RB: T-DNA borders.