Methylmercury is a common environmental contaminant in the Great Lakes region of the United States. The glutathione pathway (GSH) involves antioxidant signaling and is responsible for the efflux of toxins such as methylmercury from cells. The object of this study was to verify and characterize mutations in the GSH-associated gene gstp2 in zebrafish. We crossed prospective mutant zebrafish with wildtype zebrafish in an effort to create heterozygous and homozygous mutant zebrafish. To identify mutant zebrafish, we used PCR and restriction enzyme reagents that assess the gstp2 DNA sequence targeted by CRISPR-Cas9 reagents for mutation. Gel electrophoresis was used to visualize the results. This approach was able to identify the presence of mutated gstp2 DNA sequences at the target site. Creation of a homozygous mutant gstp2 zebrafish line will allow for experiments on the influence of gstp2 in embryonic development and developmental toxicity.
