Fig. 1.

Effects of the length of homologous sequences and presence of bait sequence on the efficiencies of targeted gene integration. (a) Schematics of each donor plasmid for evaluating the effects of homologous sequences and the bait sequence. The pGbait-acta1_500 bp-mAG plasmid contains long homology arms (Long HAs) and Gbait. The pNoBait-acta1_500 bp-mAG plasmid is the plasmid that removed the Gbait from pGbait-acta1_500 bp-mAG plasmid. The pGbait-acta1_40 bp-mAG plasmid contains 40 bp short homology arms (Short HAs) and Gbait. The pGbait-acta1_20 bp-mAG plasmid contains 20 bp short homology arms (Short HAs) and Gbait. (b) Embryos at 4 days post fertilization (dpf). Embryos injected with each plasmid, Cas9 RNA (100 ng/μl) with or without Gbait-sgRNA were categorized into the following two groups; “No GFP” for embryos without green fluorescence and “GFP+” for embryos with fluorescence in the skeletal muscle, respectively. BF: bright field, GFP: GFP fluorescent image