Figure 6.

NHEJ-mediated gene replacement in plants. A, Structure of the target gene region and outcome of gfp replacement events in Arabidopsis (A15) and tobacco (T19 and T14) plants. The locations and names of the primers used for PCR amplification of the replacement events are shown. F, Filler DNA; gfp, GFP-encoding gene; hpt, hygromycin resistance-encoding gene; P, dual CaMV 35S promoter; QQR, QQR ZFN recognition site; QQR*, altered QQR ZFN recognition site; T, CaMV 35S terminator. B, PCR analysis of wild-type (WT), parental (PR), and gfp replacement events in A15, T19, and T14 plants. M, DNA ladder. C, Sequencing data of gfp replacement events. QQR ZFN-binding sites are in blue, QQR ZFN-binding sites derived from donor DNA are underlined, gfp and hpt sequences are in green and orange, respectively, and filler DNA sequence is in purple. [See online article for color version of this figure.]