22-P001 FLEET: A new tool for gain of function and cell lineage analysis in zebrafish

Over the last 15 years, the use of the Gal4/UAS system for inducing gain-of-function clones in Drosophila melanogaster (Brand and Perrimon, 1993) has been one of the most powerful tools to uncover gene functions and to trace the fate of different cell populations during development. The use of such technology would greatly enable analysis of vertebrate development in a model system such as zebrafish, which has major advantages for invivo imaging of cells but has less well developed genetic tools. Based on the Gal4/UAS technology, and adapting it by use of strategies known to be more efficient in zebrafish, we have developed FLEET (Fish Lineage and Ectopic Expression Tool) which enables the generation of GOF clones and cell lineage tracing in zebrafish with the same DNA construct. We will show how, through the combined use of the Gal4/UAS, Cre/lox and Tol2 systems, we are able to mosaically overexpress different genes and follow the fate of a given cell population through different developmental stages.