Fig. 2

CRISPR/Cas9-based OsIMPα1b gene editing and removal of transgenes by genetic segregation. a OsIMPα1b structural features along with the Cas9-targeting sites based on the T-DNA construction of an sgRNA motif linked to the Cas9-recognizable PAM consisting of NGG (CGG here). b Nucleotide changes in the targeting sequences of the OsIMPα1b-edited (OsIMPα1b-ED) T0 lines OsIMPα1b-ED#6 and OsIMPα1b-ED#17, both of which were further investigated. c Partial fragment sequences of OsIMPα1b in WT plants (rice variety Nipponbare) and T1 progenies of the OsIMPα1b-ED#6 and OsIMPα1b-ED#17 mutants. d Genetic segregation in which the transgenes were removed during the generation of T1 and T2 progenies of both mutants tested in comparison with the WT plants. The transgenes were analyzed via PCR using genomic DNA extracted from the corresponding plant templates, and the housekeeping gene OsActin was used as a reference. Each electrophoretic agarose image is representative of 3 experimental replicates