Fig 7. Simplified procedure for generating genome edited tansgene-free mutants by using FT expression cassette-containing CRISPR/Cas9 construct.

Plants can be transformed and transgenic plants can be selected in a way similar to that for other constructs. Within the T1 transgenic plants, select plants with early-bolting phenotype, and sequence to examine genome editing status. Keep only genome edited plants. In the T2 plants germinated from seeds of the individual T1 plants, keep only those that bolt normally, and sequence to identify genome edited plants. These plants are genome edited transgene-free mutants. If necessary, confirm the transgene-free status by PCR amplification of Cas9 fragment.