Dong, Jie (Janet)

The goal of my work is to generate genetically engineered zebrafish in which transgenes can be easily deleted, replaced or mobilized using Cre recombinase and/or Tc1-like transposase. First, a construct will be generated that is capable of inserting the appropriate lox sites into the genome of zebrafish, providing a site where transgenes can be easily replaced and deleted. Our model transgene consists of the carp Beta-actin promoter and the marker gene gfp.This transgene is flanked by heterospecific lox sites(loxP and loxY) as well as Tc1 inverted repeats (TcIR). An additional loxP site is incorporated into this vector in order to facilitate the deletion of the gfp gene. Once this transgene has been integrated into the fish genome, microinjection of Cre recombinase will result in single copy inserts in which the gfp gene has been removed due to simple deletion between identical lox sites. The gfp gene will then be re-introduced into the exact same locus by microinjection with replacement plasmid carrying the gfp gene flanked by the same lox sites.