Figure 5.

SpCas9‐induced mutations in independent barley T1 lines. (a–b) Homozygous mutations in T1 plants containing the HvU3:sgRNA _ZmUbi:Cas9 and the OsU3:sgRNA _ZmUbi:Cas9 construct, respectively. (c) Heterozygous mutants have characteristic double peaks in the chromatogram, for example starting 4 bp upstream of the PAM sequence (grey). (d) Mutation patterns of heterozygous mutants determined after sequencing using specific primers (Table S1) from both directions. (e) Examples for mutations in homozygous bi‐allelic T1 lines. The PAM (CGG) sequence is highlighted in grey, the 20 bp target region in HvMORC1 is underlined, and insertions are marked in bold.