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Fig. 1. Expression of the Pax6 reporter co-injected with I-SceI meganuclease into X. tropicalis embryos. (A) A reporter construct (IS-Pax6GFP) introduced into embryos shown in panels B–G, and I. The Pax6 promoter-GFP cassette is flanked at both ends by two I-Sce1 recognition sites. (B–D) GFP fluorescence observed in embryos co-injected with IS-Pax6GFP and I-SceI meganuclease (B, stage 20; C, stage 25; D, stage 41). (E) In situ hybridization analysis of GFP expression in a stage 26 embryo co-injected with IS-Pax6GFP and I-SceI meganuclease. The smooth expression in the eye and brain tissues and the spotty ectopic expression in ventral cells are indicated by arrows and arrowhead, respectively. (F,G) Putative half transgenic embryos showing GFP expression only in one side (F, GFP fluorescence at stage 41; G, in situ hybridization with GFP probe at stage 26). (H) GFP expression in stage 41 transgenic embryo generated by the REMI method using the same Pax6 promoter-GFP cassette. (I) Mosaic GFP expression in stage 41 embryo injected without I-SceI. |
