| Humans cannot regenerate their kidneys, whereas zebrafish can regrow new kidney tissues after injury. To study this process, we set out to generate a transgenic zebrafish line in which a specific part of the kidney, the proximal convoluted tubule (PCT) is labeled with GFP. A DNA construct carrying the PCT-specific slc20a1a gene promoter was chosen for this process. First, we modified it so that it would carry the Tol2 cassette for transposon-mediated transgenesis. Then, a switchable reporter cassette (attP1-GFP-2A-CreER-attP2) was inserted into the DNA construct so that the slc20a1a promoter would control expression of the GFP reporter. The attP1/attP2 recombination sites flanking the reporter cassette would allow for future exchange of a different cassette, such as a gene that produces a toxic protein, for PCT-specific ablation. Our data suggest that the final DNA construct was successfully generated and it is ready for injection into zebrafish embryos for transgenesis upon sequencing. |
