Fig. 4

Isolation of KJFTSY knockout lines using DNA-free genome editing. a Structure of the KJFTSY gene and gRNA-targeting site. Gray boxes represent exons; yellow triangle represents the gRNA-targeting site. b Color selection of a KJFTSY knockout cell. After electroporation of Cas9-gRNA RNP, electroporated cells were spread on a plate to obtain single colonies. An enlarged view of a KJFTSY knockout colony is also shown. The photograph was taken after a 7-day incubation in dark followed by a 14-day incubation in light. c The gRNA target sequence in the wild-type KJFTSY gene and the corresponding sequences in four FTSY knockout lines. Red letters indicate inserted nucleotides. The PAM sequence is shown in green. d Partial amino acid sequence of wild-type KJFTSY and those of KJFTSY knockdown lines. The red letters indicate amino acid sequences generated by frameshift mutations