Creation of targeted mutations in the Zebrafish using zinc finger nucleases

Zinc finger nucleases (ZFNs) have been shown to be effective for gene targeting in the Zebrafish. ZFNs are engineered that recognize sequences in Zebrafish genes. Co-injection of mRNAs encoding these ZFNs into one-cell-stage Zebrafish embryos lead to mutagenic lesions at the target site that are transmitted through the germ line with high frequency. This project was created to identify lesions created by ZFNs in pools of Zebrafish embryos for rapid genotyping of ZFN injected embryos, to negate the need for cloning and re-sequencing.