Molecular characterization of the tagged CESA10 locus and the mEGFP-CSLD6-TEN locus. (A) Diagram illustrates the result of HDR mediated insertion of mEGFP sequences from the homology repair plasmid (bottom) into the CESA10 genomic locus (top). Exons (first 7 shown) are indicated by pink boxes and the cloned promoter is indicated by cyan boxes. Thin lines indicate intronic regions. The inserted mEGFP sequence is denoted by a thick green box. Small arrows above the diagrams represent primers used for genotyping. (B) PCR products obtained with primer pairs using template DNA isolated from the indicated moss lines were separated on an agarose gel and stained with ethidium bromide. The asterisk indicates the line chosen for imaging after sequencing the PCR product. Molecular weight is indicated in kb. Predicted sizes for correct products are indicated below the gel. (C) Sequencing of a PCR product (Table S2) amplified from a plant transformed with PS4 (yellow bar) cloned into pMH-Cas9 together with double stranded TEN-oligo (Table S2) reveals CRISPR-Cas9 mediated editing resulting in the three designed silent mutations and the G2599A mutation resulting in D867N. Source data are available for this figure: SourceData FS4.