Transposon-Based Platform for Insertional Mutagenesis and Genome Engineering Abstract: The fastest, most comprehensive and cost-effective way to study human gene function is through their counterparts in model systems such as zebrafish. Two key characteristics of any gene are expression pattern and loss of function phenotype. We propose to develop tools that for the first time in a vertebrate simultaneously assay both of these characteristics. Our vectors will report gene expression patterns using Gal4/UAS while conditionally mutating genes in zebrafish. Gal4/UAS will enable investigators to use our gene traps as driver lines to express other proteins of interest in specific tissues. Gene trap cassettes will be flanked by a combination of site-specific recombinase recognition sites for regulation of mutagenicity. During the process of improving and testing our vectors we will generate, propagate and make available 150 zebrafish gene trap lines. To complement these gene trap lines, we will produce several lines with tissue-specific expression of Cre recombinase for spatial regulation of mutagenicity and use them to revert integrations into genes essential for viability. These tissue-specific Cre lines will be also useful for genetic lineage tracing. Since our gene traps will contain recognition sites for site-specific recombinases, the 120 gene trap lines will constitute a library of recombinase recognition sites distributed throughout the zebrafish genome. This will allow us to carry out proof of principle experiments for use of site-specific recombinases to generate large inversions and deletions, and enable other researchers to generate zebrafish """"""""designer chromosomes"""""""" of interest.

Public Health Relevance

Genetics plays a substantial role in many human diseases. This project will develop tools to greatly enhance our ability to study gene function in a vertebrate model system- the zebrafish.

National Institute of Health (NIH)
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-BDA-L (50))
Program Officer
Coulombe, James N
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Temple University
Schools of Arts and Sciences
United States
Zip Code
Burg, Leonard; Palmer, Nicholas; Kikhi, Khrievono et al. (2018) Conditional mutagenesis by oligonucleotide-mediated integration of loxP sites in zebrafish. PLoS Genet 14:e1007754
Burg, Leonard; Zhang, Karen; Bonawitz, Tristan et al. (2016) Internal epitope tagging informed by relative lack of sequence conservation. Sci Rep 6:36986
Seiler, Christoph; Gebhart, Nichole; Zhang, Yong et al. (2015) Mutagenesis Screen Identifies agtpbp1 and eps15L1 as Essential for T lymphocyte Development in Zebrafish. PLoS One 10:e0131908
Craig, Michael P; Grajevskaja, Viktorija; Liao, Hsin-Kai et al. (2015) Etv2 and fli1b function together as key regulators of vasculogenesis and angiogenesis. Arterioscler Thromb Vasc Biol 35:865-76
Roberts, Jennifer Anne; Miguel-Escalada, Irene; Slovik, Katherine Joan et al. (2014) Targeted transgene integration overcomes variability of position effects in zebrafish. Development 141:715-24
Burg, Leonard; Gill, Ryan; Balciuniene, Jorune et al. (2014) SideRack: a cost-effective addition to commercial zebrafish housing systems. Zebrafish 11:167-72
Malafoglia, Valentina; Colasanti, Marco; Raffaeli, William et al. (2014) Extreme thermal noxious stimuli induce pain responses in zebrafish larvae. J Cell Physiol 229:300-8
Grajevskaja, Viktorija; Balciuniene, Jorune; Balciunas, Darius (2013) Chicken ?-globin insulators fail to shield the nkx2.5 promoter from integration site effects in zebrafish. Mol Genet Genomics 288:717-25
Balciuniene, Jorune; Balciunas, Darius (2013) Gene trapping using gal4 in zebrafish. J Vis Exp :e50113
Balciuniene, Jorune; Nagelberg, Danielle; Walsh, Kathleen T et al. (2013) Efficient disruption of Zebrafish genes using a Gal4-containing gene trap. BMC Genomics 14:619