Figure 7.

Homology‐directed repair with a DNA donor homology plasmid that contains a protospacer targeting sequence. (a) A partial gene model representing the 5′ region of Pp3c16_8300. The black arrowhead represents the desired mRuby2 insertion site. The DNA sequence of the 3′ homology fragment is shown within the blue arrow, with the protospacer binding sequence shown in red and the PAM shown in a red box. Plants co‐transformed with the DNA donor template and the Cas9 plasmid were unable to integrate the mRuby2 sequence into the locus (right). (b) The DNA sequence of the 3′ homology fragment after introduction of a silent mutation within the protospacer binding sequence (bold, asterisks). Mutagenesis allows mRuby2 integration upon co‐transformation with the Cas9 plasmid (right). Sequenced plants are indicated with bold and underlined numbers