Fig. 6

Transgene mirrors the endogenous expression of the target gene. (A-F) Lateral view of 2dpf sox2-2a-sfGFP^stl84;gfap-2a-tdTomato^stl85 double homozygous knock-in embryos. All images were taken with the same exposure time. (A-C) sfGFP expression. (D-F) tdTomato expression. (A,D) Control embryo injected with 100pg of cas9 RNA. (B,E) The embryo co-injected with 100pg of cas9 RNA and 10pg of gRNA targeting sox2. (C,F) The embryo co-injected with 100pg of cas9 RNA and 10pg of gRNA targeting gfap. Scale bar: 200µm. (G,H) Detection of target-specific mutations by T7EI assays. Control, embryo injected with 100 pg of cas9 RNA; 1-5, individual embryos co-injected with 100pg of cas9 RNA and 10pg of gRNA targeting sox2 (G) or gfap (H). (G) The PCR products contain guide RNA target site of sox2. (H) The PCR products contain guide RNA target site of gfap.