Cre/lox-regulated transgenic zebrafish model with conditional myc-induced T cell acute lymphoblastic leukemia

Proc Natl Acad Sci U S A. 2005 Apr 26;102(17):6068-73. doi: 10.1073/pnas.0408708102. Epub 2005 Apr 12.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • DNA-Binding Proteins / genetics
  • Extracellular Matrix Proteins / metabolism
  • Genes, myc*
  • Genetic Markers
  • Green Fluorescent Proteins / genetics*
  • Humans
  • Integrases / metabolism*
  • Leukemia-Lymphoma, Adult T-Cell / genetics*
  • Mice
  • Molecular Sequence Data
  • Nuclear Proteins
  • Protein-Lysine 6-Oxidase / metabolism
  • Zebrafish / genetics

Substances

  • DNA-Binding Proteins
  • Extracellular Matrix Proteins
  • Genetic Markers
  • Nuclear Proteins
  • RAG2 protein, human
  • Rag2 protein, mouse
  • V(D)J recombination activating protein 2
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Lox protein, mouse
  • Protein-Lysine 6-Oxidase
  • Cre recombinase
  • Integrases

Associated data

  • PIR/AF398514