Fig. 1
From: Gene targeting and transgene stacking using intra genomic homologous recombination in plants
Basic design of constructs used for gene targeting via intra genomic homologous recombination. a Target construct contains a generation 1 (Gen 1) stack and a selection marker A flanked by unique homology sequences (HR1 and 2). A designed nuclease 1 binding site is inserted between selection marker A and HR2 sequence. b The donor construct contains selection marker B, downstream HR3 sequence, generation 2 (Gen 2) stack, and HR1 and 2 homology sequences matching the target. A designed nuclease 2 binding site is inserted between the selection marker and HR3 sequence for future targeting. The donor is flanked by a designed nuclease 1 binding site on each end. c The designed nuclease construct contains designed nuclease 1 coding sequence driven by appropriate promoter. d Target locus containing functional selection marker B gene and generation 2 (Gen 2) stack precisely inserted after gene targeting