Transgenic reporter zebrafish

Perz-Edwards, A., N.L. Hardison and E. Linney. Retinoic acid-mediated gene expression in transgenic reporter zebrafish. Dev. Biol. 229 89-101, 2001. 

The protocol described here spans over eight experimental days. First, transgenic embryos are generated by crossing wild-type and transgenic pckl reporter zebrafish. At 4 dpf, the transgenic larvae are distributed in 96-well plates and treated with small molecules. After a 48-h incubation time, the bioluminescence signal is measured and serves as a readout for pckl promoter activity. 

At 4 dpf verify expression of the red fluorescence marker in the eye that indicates transgenic Tp (pckl Lm2) reporter zebrafish using a fluorescence stereoscope. 

Schreurs, R., et al.. Estrogenic activity of UV filters determined by an in vitro reporter gene assay and an in vivo transgenic zebrafish assay. Arch. Toxicol, 76, 257-61, 2002. 

The first transgene to be delivered into medaka embryos was the 6-crystalline gene of chicken (10) resulting in transient mosaic expression of the transgene. In 1988, Stuart McMurray, and Westerfield reported the first successful germline transmission and subsequent expression of a transgene on microinjection in a teleost genetic model system (zebrafish (12)). 

Set up 10-30 crosses of homozygous T f(pckl Luc2) with wild-type zebrafish to generate 1,000-3,000 embryos that are hemizygous for the pckl reporter transgene see Note 4). For each cross use one male and one to two females. 

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