Figure 2 | Scientific Reports

Figure 2

From: True gene-targeting events by CRISPR/Cas-induced DSB repair of the PPO locus with an ectopically integrated repair template

Figure 2

Analysis of parental lines. (A) T2 transformants containing the LSL-PPO repair template and the Cas9-PPO nuclease gene were germinated on kanamycin and phosphinothricin (P1-P8) and the transformants containing the 35S-REP gene on hygromycin (R119 and R124). The size bar is 1 cm. (B) Footprint analysis of plant lines P1-P8. Genomic DNA was pre-digested with FauI or not pre-digested and the PPO target site was amplified and an aliquot was separated on a 1.5% agarose gel. The PCR products were digested with FauI or a combination of FauI and KpnI and separated on 1.5% agarose gels. A negative control not containing template DNA (−) was included in the PCR reaction. M is the 1 kb DNA marker. The full-size gels are presented in Fig. S4. (C) Sequences of cloned target sites of P3 and P8. The sgRNA sequence is in yellow, PAM sequence in grey, insertions are in blue. Deletions are indicated by dashes and repair via microhomology in pink.

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