Transgenic zebra fish embryo model for hematopoiesis and lymphoproliferative disorders

Abstract

A transgenic zebrafish animal model for the study of haemopoietic cell differentiation, control, and screening of therapeutic agents.

1 . An animal model for lymphocyte development and leukemia, comprising a transgenic zebrafish expressing a heterologous Ikaros protein.  
     
     
         2 . The animal model of  claim 1 , wherein the Ikaros protein is a non-DNA binding form of Ikaros.  
     
     
         3 . The animal model of  claim 2 , wherein the Ikaros protein lacks at least one N-terminal zinc finger domain as compared with DNA-binding forms of Ikaros.  
     
     
         4 . The animal model of  claim 3 , wherein the Ikaros protein is one or more of Ik-4, Ik-5, Ik-6, Ik-7, and Ik-8.  
     
     
         5 . The animal model of  claim 3 , wherein the Ikaros protein is a deletion mutant lacking the following Ikaros amino acid sequence: KSSMPQKFLG [SEQ ID NO: 5].  
     
     
         6 . The animal model of  claim 3 , wherein the Ikaros protein contains an insertion of the following amino acid sequence: VTVGADDFRDFHAIIPKSFSR [SEQ ID NO: 6].  
     
     
         7 . An assay method for screening potential therapeutic agents useful for treating or preventing hematopoietic disorders, the assay comprising contacting a transgenic zebrafish embryo with a potential therapeutic agent, the transgenic zebrafish embryo expressing a non-DNA binding form of Ikaros, and correlating improved lymphohematopoiesis versus a non-treated control with an effective therapeutic agent.  
     
     
         8 . The assay of  claim 7 , wherein the non-DNA binding form of Ikaros protein lacks at least one N-terminal zinc finger domain as compared with DNA-binding forms of Ikaros.  
     
     
         9 . The assay of  claim 7 , wherein the Ikaros protein is one or more of Ik-4, Ik-5, Ik-6, Ik-7, and Ik-8.  
     
     
         10 . The assay of  claim 7 , wherein the Ikaros protein is a deletion mutant lacking the following Ikaros amino acid sequence: KSSMPQKFLG [SEQ ID NO: 5] 
     
     
         11 . The assay of  claim 7 , wherein the Ikaros protein contains an insertion of the following amino acid sequence: VTVGADDFRDFHAIIPKSFSR [SEQ ID NO: 6].  
     
     
         12 . The assay of  claim 7 , wherein said improved lymphohematopoiesis is analyzed in the developing zebrafish embryo.  
     
     
         13 . The assay of  claim 12 , wherein said improved lymphohematopoiesis comprises improved oligochromemia in the circulating blood cells of the animal model.  
     
     
         14 . The assay of  claim 7 , wherein said improved lymphohematopoiesis is analyzed in adult zebrafish.  
     
     
         15 . The assay of  claim 7 , wherein said improved lymphohematopoiesis comprises improved cellularity and celllular composition of adult zebrafish kidney imprints.  
     
     
         16 . The assay of  claim 7 , wherein said improved lymphohematopoiesis comprises one or more of lessened multilineage hematopoiesis, reduced erythroid hyperplasia, and reduced numbers of lymphoblasts as compared with control animals.