FIGURE 3.

Analysis of genome editing at the mCherry locus. Cells of the SC6 line were transformed with pFGC-Cas9-sgRNA1-2-3 targeting mCherry. Transformants appeared after 3 weeks on bialaphos selection medium and were transferred twice on new selection medium. (A) Picture of the calli under visible light (left) and fluorescence of mCherry (right). (B) Close-up of four calli: lines 15 (homogenous mCherry fluorescence), 18 (no mCherry fluorescence) 6 and 20 (heterogenous mCherry fluorescence). (C) Genome editing in transformed calli was monitored by PCR amplification of mCherry. Deletion between two target sites occurred in seven out of 20 transformants (lines 1, 2, 8, 9, 17, 18, 19). pPZP: amplification of mCherry directly from the plasmid pPZP-mCherry. (D) Genotyping of nine transformants by RFLP analysis. The PCR fragments from lines 1 to 9 displayed in (C) were subjected to digestion by the indicated restriction enzymes.