Methylmercury (MeHg) toxicity is a notable public health concern in the Great Lakes region. The glutathione pathway (GSH) involves antioxidant signaling and is responsible for the removal of toxins like MeHg from cells. There are many genes involved in the GSH pathway; one of these genes is gclm. Previous lab work has developed a mutant zebrafish line by creating a germline mutation using CRISPR-Cas9 reagents that contains a loss-of-function deletion in the gclm gene. The objective of this study was to create additional gclm mutant fish for investigating developmental MeHg toxicity. Zebrafish of different gclm genotypes were bred to produce heterozygous and homozygous mutant zebrafish. To identify mutant zebrafish, we used PCR and restriction enzyme reagents that target the DNA mutation site in the gclm gene. Results were visualized using gel electrophoresis. The creation of a mutant gclm zebrafish line will allow for future experiments on the influence of gclm in embryonic development and developmental toxicity.
