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Fig. 1
In vivo detection system for targeted integration of reporter constructs in zebrafish. (A) Schematic of PhiC31 targeted integration system. Transgenic embryos containing an attP docking site have a green lens due to the gamma-crystalline promoter driving a GFP reporter. ITR labels recognition sequences of either Tol2 or Sleeping beauty transposases used in generating the recipient transgenic lines with docking site. Injection of a circular plasmid with attB and a red reporter (targeting vector) into recipient line eggs leads to eye colour change upon PhiC31-targeted integration in larvae. (B) Detection of eye colour change upon PhiC31-mediated integration in the transgenic recipient Tg(Xla.crygc:attP-GFP)uobL6 line. Top row shows a transgenic larva with PhiC31-mediated recombination of the attB-mCherry cassette into the attP-GFP docking site. Bottom row shows transgenic sibling from the recipient line without targeted integration. Side views on cranial end of 5 dpf larvae with anterior to the left. Scale bar: 100 µm. Arrows indicate reporter expression in the lens. (C) Sequence of the tpl102 recipient docking site with germline integration of targeting construct tnnT2:attB-mRFP. Sequence shows the attR site in the crygc:attR-Red recombination product. The three lower case nucleotides denote the recombination site.