Etv2 and uses thereof

Abstract

Described herein is a method for producing a chimeric non-human animal expressing a human ETV2 gene comprising: a) generating an ETV2 null non-human animal cell, wherein both copies of the non-human ETV2 gene carry a mutation that prevents production of functional ETV2 protein in said non-human animal; b) creating an ETV2 null non-human blastocyst by somatic cell nuclear transfer comprising fusing a nucleus from said ETV2 null non-human animal cell of a) into an enucleated non-human oocyte and activating said oocyte to divide so as to form an ETV2 null non-human blastocyst; c) introducing human stem cells into the ETV2 null non-human blastocyst of b); and d) implanting said blastocyst from c) into a pseudopregnant surrogate non-human animal to generate a chimeric non-human animal expressing human ETV2.

1 . A non-human animal cell or blastocyst wherein the genome carries a mutation in both alleles of the ETV2 gene such that the non-human animal cell or blastocyst lacks functional ETV2 protein. 
     
     
         2 . The non-human animal or blastocyst of  claim 1 , wherein the mutation is a deletion of the ETV2 gene. 
     
     
         3 . The non-human animal cell or blastocyst of  claim 1 , wherein the non-human animal cell or blastocyst is a porcine, bovine, equine or goat. 
     
     
         4 . A chimeric non-human animal or blastocyst expressing human ETV2 and lacking expression of said non-human animal ETV2. 
     
     
         5 . The chimeric non-human animal of  claim 4 , wherein the non-human animal expresses human blood cells selected from the group consisting of white blood cells, red blood cells, platelets or a combination thereof. 
     
     
         6 . The chimeric non-human animal of  claim 4 , wherein the chimeric non-human animal expresses human endothelium. 
     
     
         7 . The chimeric non-human animal of  claim 4 , wherein the non-human animal is a porcine, bovine, equine or goat. 
     
     
         8 . A method for producing a chimeric non-human animal expressing a human ETV2 gene comprising:
 a) generating an ETV2 null non-human animal cell, wherein both copies of the non-human ETV2 gene carry a mutation that prevents production of functional ETV2 protein in said non-human animal;   b) creating an ETV2 null non-human blastocyst by somatic cell nuclear transfer comprising fusing a nucleus from said ETV2 null non-human animal cell of a) into an enucleated non-human oocyte and activating said oocyte to divide so as to form an ETV2 null non-human blastocyst;   c) introducing human stem cells into the ETV2 null non-human blastocyst of b); and   d) implanting said blastocyst from c) into a pseudopregnant surrogate non-human animal to generate a chimeric non-human animal expressing human ETV2.   
     
     
         9 . A method of producing human and/or humanized blood cells or vessels in a non-human animal comprising:
 a) generating an ETV2 null non-human animal cell, wherein both alleles of the non-human ETV2 gene carry a mutation that prevents production of functional ETV2 protein;   b) creating an ETV2 null non-human blastocyst or morula by somatic cell nuclear transfer comprising fusing a nucleus from said ETV2 null non-human animal cell of a) into an enucleated non-human oocyte and activating said oocyte to divide so as to form an ETV2 null non-human blastocyst;   c) introducing human donor stem cells into the ETV2 null non-human blastocyst of b); and   d) implanting said blastocyst or morula from c) into a pseudopregnant surrogate non-human animal so as to generate a non-human animal expressing human and/or humanized blood cells or vessels.   
     
     
         10 . The method of  claim 8 , wherein the non-human animal is a porcine, bovine, equine or goat. 
     
     
         11 . The method of  claim 8 , wherein the human donor stem cell is a tissue specific stem cell, pluripotent stem cell, multipotent adult stem cell, induced pluripotent stem cell or umbilical cord blood stem cell (UCBSC). 
     
     
         12 . The method of  claim 9 , wherein the donor providing the stem cells is the recipient of the humanized tissue or organ produced. 
     
     
         13 . The method of  claim 9 , wherein the human induced pluripotent cell is formed from a fibroblast cell. 
     
     
         14 . A non-human animal produced by the method of  claim 8 . 
     
     
         15 . A progeny non-human animal produced from the mating of two non-human animals generated from the method of  claim 8 , wherein said progeny non-human animal expresses human ETV2, wherein the genome of said progeny non-human carries a homozygous deletion of said non-human animal ETV2 gene. 
     
     
         16 . The progeny non-human animal of  claim 15 , wherein the non-human animal is a porcine, bovine, equine or goat. 
     
     
         17 . The method of  claim 9 , wherein the blood cells are human-induced pluripotent or human umbilical cord blood stem cell-derived white blood cells, red blood cells, and/or platelets. 
     
     
         18 . Chimeric blood cells made by a process comprising:
 a) generating an ETV2 null non-human animal cell, wherein both alleles of the non-human ETV2 gene carry a mutation that prevents production of functional ETV2 protein;   b) creating an ETV2 null non-human blastocyst or morula by somatic cell nuclear transfer comprising fusing a nucleus from said ETV2 null non-human animal cell of a) into an enucleated non-human oocyte and activating said oocyte to divide so as to form an ETV2 null non-human blastocyst;   c) introducing human donor stem cells into the ETV2 null non-human blastocyst of b); and   d) implanting said blastocyst or morula from c) into a pseudopregnant surrogate non-human animal so as to generate a non-human animal expressing chimeric blood cells.   
     
     
         19 . A blood cell, wherein the cell expresses the human ETV2 gene and is derived from an implanted non-human blastocyst or morula comprising human donor cells. 
     
     
         20 . The chimeric blood cells of  claim 19 , wherein the non-human animal is swine.