Fig. 4

Generation of FAE1 knockout using CRISPR/Cas9 system in pCam5 transgenic line. a Fatty acid synthesis by FAE1 in transgenic Arabidopsis expressing RcFAH12. A solid arrow indicates strong flux, dashed arrow indicates weak flux. b CRISPR/Cas9 vector map for knockout of FAE1 gene in Arabidopsis. HygR, hygromycin resistance gene is driven by CaMV promoter; Cas9 gene is driven by Egg cell-specific promoter; sgRNA is driven by U6 promoter; LB/RB, left border, right border; CaMV P, Cauliflower mosaic virus gene promoter; EC2.1 P, Egg cell-specific promoter; U6 26P, U6 29P, U6 promoter. c Schematic diagram of design for sgRNAs in FAE1 gene. Two single guide RNA (sgRNA1, 2) positions for FAE1 gene editing. To identify the deletion pattern of the FAE1 gene, two primers are designed. d Sanger sequencing result of four independent pCam5-atfae1 lines with addition or deletion of FAE1 gene. Red color indicates PAM(NGG) sequence and blue color indicates mismatch sequence compared to wild type