Genetically Modified Non-Human Animals And Methods Of Use Thereof

Abstract

The invention relates generally to genetically modified non-human animals expressing human polypeptides and their methods of use.

1 . A genetically modified mouse comprising in its genome:
 a recombination activating gene 2 (Rag-2) gene knock-out;   an IL2 receptor gamma chain (IL2rg) gene knock-out;   a replacement of a mouse M-CSF gene with a nucleic acid encoding a human M-CSF polypeptide, wherein the nucleic acid encoding the human M-CSF polypeptide is operably linked to a M-CSF promoter at a mouse M-CSF gene locus;   a replacement of a mouse IL-3 gene with a nucleic acid encoding a human IL-3 polypeptide, wherein the nucleic acid encoding the human IL-3 polypeptide is operably linked to a IL-3 promoter at a mouse IL-3 gene locus;   a replacement of a mouse GM-CSF gene with a nucleic acid encoding a human GM-CSF polypeptide, wherein the nucleic acid encoding the human GM-CSF polypeptide is operably linked to a GM-CSF promoter at a mouse GM-CSF gene locus;   a replacement of a mouse-SIRPA gene with a nucleic acid encoding a fusion protein comprising a biologically active fragment of a full length human SIRPA polypeptide, wherein the nucleic acid encoding the fusion protein is operably linked to a SIRPA promoter; and   a replacement of a mouse TPO gene with a nucleic acid encoding a human TPO polypeptide, wherein the nucleic acid encoding the human TPO polypeptide is operably linked to a TPO promoter at a mouse TPO gene locus, wherein the mouse expresses the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the fusion protein, and the human TPO polypeptide.   
     
     
         2 .- 8 . (canceled) 
     
     
         9 . The genetically modified mouse of  claim 1 , further comprising human hematopoietic cells. 
     
     
         10 . The genetically modified mouse of  claim 1 , further comprising a human cancer cell. 
     
     
         11 . The genetically modified mouse of  claim 10 , wherein the human cancer cell is a leukemia cell or a melanoma cell. 
     
     
         12 . A method of hematopoietic stem and progenitor cell (HSPC) engraftment in a genetically modified mouse, the method comprising the step of: administering at least one HSPC to the genetically modified mouse, wherein the genetically modified mouse comprises in its genome:
 a recombination activating gene 2 (Rag-2) gene knock-out;   an IL2 receptor gamma chain (IL2rg) gene knock-out;   a replacement of a mouse M-CSF gene with a nucleic acid encoding a human M-CSF polypeptide, wherein the nucleic acid encoding the human M-CSF polypeptide is operably linked to a M-CSF promoter at a mouse M-CSF gene locus;   a replacement of a mouse IL-3 gene with a nucleic acid encoding a human IL-3 polypeptide, wherein the nucleic acid encoding the human IL-3 polypeptide is operably linked to a IL-3 promoter at a mouse IL-3 gene locus;   a replacement of a mouse GM-CSF gene with a nucleic acid encoding a human GM-CSF polypeptide, wherein the nucleic acid encoding the human GM-CSF polypeptide is operably linked to a GM-CSF promoter at a mouse GM-CSF gene locus;   a replacement of a mouse-SIRPA gene with a nucleic acid encoding a fusion protein comprising a biologically active fragment of a full length human SIRPA polypeptide, wherein the nucleic acid encoding the fusion protein is operably linked to a SIRPA promoter; and   a replacement of a mouse TPO gene with a nucleic acid encoding a human TPO polypeptide, wherein the nucleic acid encoding the human TPO polypeptide is operably linked to a TPO promoter at a mouse TPO gene locus, wherein the mouse expresses the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the fusion protein, and the human TPO polypeptide.   
     
     
         13 .- 18 . (canceled) 
     
     
         19 . The method of  claim 12 , wherein the genetically modified mouse comprises a human cancer cell. 
     
     
         20 . The method of  claim 19 , wherein the human cancer cell is a leukemia cell or a melanoma cell. 
     
     
         21 .- 27 . (canceled) 
     
     
         28 . The method of  claim 12 , wherein the administering step is not preceded by preconditioning irradiation of the genetically modified mouse or administration of a radiomimetic drug to the genetically modified mouse. 
     
     
         29 . The method of  claim 12 , further comprising administering a human cancer cell to the genetically modified mouse. 
     
     
         30 . The method of  claim 29 , wherein the human cancer cell is a leukemia cell or a melanoma cell. 
     
     
         31 . The method of  claim 29 , wherein the at least one HSPC and the human cancer cell are both isolated from the same patient.