Transposon-mediated Insertional Mutagenesis Screen

We have conducted an insertional mutagenesis screen using a P9 gene-breaking cassette, a Tol2-based transposon vector that was developed in Dr. Stephen Ekker’s lab. We have successfully identified 45 embryonic lethal mutants, indicating that this is so far the most efficient strategy to conduct an insertional mutagenesis screen in zebrafish. We are investigating the molecular mechanisms of the screening strategy by cloning the corresponding genes for these mutants, so that this efficient technology can be widely used in the zebrafish community to facilitate the generation of mutants in every gene in the zebrafish genome.

Currently we are focusing on eight mutants that affect different steps of cardiogenesis, including the migration of cardiac progenitors to the midline to form the heart tube, the establishment of cardiac asymmetry, and the development of cardiac neural crest cells.