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. 2017 Dec 4;16(6):1125–1137. doi: 10.1111/pbi.12857

Figure 3.

Figure 3

Integration sites and subsequent segregation of the CRISPR/Cas9 transgene targeting the GmDrb2 loci. (a) WGS analysis identified three transgene integrations located on chromosomes four, thirteen and fifteen, respectively. Green arrows represent primers used to screen for null‐segregants (Table S5), the pink coloured box represents a duplicated region, the orange coloured box represents the DNA insertion, and the grey coloured rectangle represents the unknown sequence that does not return a BLASTn search result to any organism. (b) A PCR assay using transgene‐specific primers and primers spanning the genomic integration sites was used to screen for the removal of reagent transgenes by genetic segregation. No amplicons could be detected from the transgene corresponding to any of three transgene genomic locations in the WPT590‐4‐28‐5 plant. (c) The WPT590‐4‐28‐5 plant was sequence‐confirmed and found to be homozygous for the drb2a Δ7 and drb2b Δ4 mutant alleles, and termed the Gmdrb2ab double mutant.