Ongoing projects in the laboratory include:
- Characterization of gene trap lines relevant for cardiovascular development and regeneration. We currently have a collection of over 60 gene trap lines produced with our first-generation (Cre-reversible) vectors. We are switching the screen over to second-generation (fully conditional) vectors. In this set of projects, we identify mutated genes and assess their role in development and regeneration the heart, as well as fin regeneration.
- Generation and analysis of transgenic lines expressing Cre recombinase in tissue-specific and inducible manner. Mutagenicity of our gene traps is regulated by expression of Cre recombinase. This project aims to add to our collection of transgenic Cre lines to enable manipulation of gene trap alleles in adult zebrafish in different cell types participating in regeneration of the heart and the fin.
- Analysis of transgenic lines expressing different versions of Flp recombinase. We have generated several transgenic lines expressing two versions of Flp recombinase, eFLP and Flpo. This project evaluates the potential of these recombinases to manipulate our gene traps, as well as their ability to induce chromosomal aberrations (see below).
- Development of Gal4/UAS system for high-fidelity expression of transgenes. Our gene traps use Gal4-VP16 as the primary reporter, which makes our gene trap lines suitable for expression of other transgenes under the control of Gal4 UAS. The most significant drawback of Gal4/UAS system is variegation. This project uses several complementary approaches to improve the fidelity and predictability of transgene expression from Gal4 UAS.
- Site-specific recombinases and I-SceI meganuclease as genome engineering tools. Our insertional mutagenesis is leaving the zebrafish genome peppered with sites for Cre, Flp and I-SceI. We are testing the potential of these enzymes to induce different classes of genome engineering events: deletions, inversions and homologous recombination.