Figure 3 | Scientific Reports

Figure 3

From: CRISPR/Cas9-mediated genome editing efficiently creates specific mutations at multiple loci using one sgRNA in Brassica napus

Figure 3

Phenotype and genotype of a BnaA6.RGA-sgRNA single mutant (T0) and a BnaRGA-sgRNA quadruple mutant (T1). (A) Morphology of BnaA6.RGA-sgRNA transgenic plants at the same age. L4 and L6: two individual T0 transgenic lines. CK: BnaA6.DA1-sgRNA (L10). Bar = 15 cm. (B) Genotype of BnaA6.RGA-sgRNA lines (L4 and L6) in the T0 generation. (C) Amino acid sequence alignment of the target sites derived from WT and two BnaA6.RGA-sgRNA transgenic lines (L4 and L6). The DELLA and TVHYNP domains are underlined. The amino acid deletion in the DELLA domain is caused by sgRNA1. The amino acid deletion in the TVHYNP domain is caused by sgRNA2. (D) Image showing the morphology of the quadruple mutant induced by BnaRGA-sgRNA (L46-T1) at the same age as the control. CK: WT plant. Bar = 15 cm. (E) Genotype of the quadruple mutant isolated from BnaRGA-sgRNA-L46 in the T1 generation. The PAM sequence is indicated with green. The sgRNA is indicated with red. The mutation sites are indictaed with blue. For the term n/m, m indicates the number of clones examined, and n indicates the number of clones showing the indicated genotype.

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