Cre/Lox-Regulated Transgenic Zebrafish Model with Conditional Myc-Induced T Cell Acute Lymphoblastic Leukemia

• Published in: Proceedings of the National Academy of Sciences - PNAS 2005-04, Vol.102 (17), p.6068-6073
• Main Authors: Langenau, David M., Feng, Hui, Berghmans, Stephane, Kanki, John P., Kutok, Jeffery L., Look, A. Thomas, Cooper, Max D.
• Format: Article
• Language: English
• Subjects: Acute T cell leukemia; Animals; Animals, Genetically Modified; Biological Sciences; Disease models; DNA-Binding Proteins - genetics; Embryos; Extracellular Matrix Proteins - metabolism; Fish; Fishing lines; Gene expression; Genes, myc; Genetic Markers; Genetics; Green Fluorescent Proteins - genetics; Humans; Integrases - metabolism; Leukemia; Leukemia-Lymphoma, Adult T-Cell - genetics; Mice; Molecular Sequence Data; Nuclear Proteins; Protein-Lysine 6-Oxidase - metabolism; RNA; T cell receptors; T lymphocytes; Transgenes; Transgenic animals; Transgenic plants; Zebrafish - genetics
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0408708102

We have created a stable transgenic rag2-EGFP-mMyc zebrafish line that develops GFP-labeled T cell acute lymphoblastic leukemia (T-ALL), allowing visualization of the onset and spread of this disease. Here, we show that leukemias from this transgenic line are highly penetrant and render animals moribund by 80.7 ± 17.6 days of life (±1 SD, range = 50-158 days). These T cell leukemias are clonally aneuploid, can be transplanted into irradiated recipient fish, and express the zebrafish orthologues of the human T-ALL oncogenes tal1/scl and lmo2, thus providing an animal model for the most prevalent molecular subgroup of human T-ALL. Because T-ALL develops very rapidly in rag2-EGFP-mMyc transgenic fish (in which "mMyc" represents mouse c-Myc), this line can only be maintained by in vitro fertilization. Thus, we have created a conditional transgene in which the EGFP-mMyc oncogene is preceded by a loxed dsRED2 gene and have generated stable rag2-loxP-dsRED2-loxP-EGFP-mMyc transgenic zebrafish lines, which have red fluorescent thymocytes and do not develop leukemia. Transgenic progeny from one of these lines can be induced to develop T-ALL by injecting Cre RNA into one-cell-stage embryos, demonstrating the utility of the Cre/lox system in the zebrafish and providing an essential step in preparing this model for chemical and genetic screens designed to identify modifiers of Myc-induced T-ALL.