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. 2017 Sep 7;8:1548. doi: 10.3389/fpls.2017.01548

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Copyright © 2017 Fan, Liu, Lyu, Wang, Yang and Zhu.

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) or licensor 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/Cas9-mediated knockout of Rfg1 in the Williams 82 (Nod–) background. (A) Gene structure of Rfg1. The exons and introns are indicated by boxes and lines, respectively. Arrow indicates the transcription direction. (B) Two targeted sites on the fourth exon. The protospacer adjacent motif (PAM) is ‘TGG’ for the site 1 and ‘AGG’ for the site 2. (C) An example showing that knockout of Rfg1 at the site 1 led to the formation of root nodules on a transgenic root (boxed). Sequence analysis revealed that this root contained two mutant alleles, one with a 3-bp deletion and another with a 10-bp deletion (indicated by arrows). (D) An example showing that knockout of Rfg1 at the site 2 also resulted in the formation of root nodules on a transgenic root (boxed). Sequence analysis of the DNA from this root identified two mutant alleles, one with a 4-bp deletion and another with a 26-bp deletion (indicated by arrows).