Non-human animals having a humanized cluster of differentiation 47 gene

Abstract

Non-human animals, and methods and compositions for making and using the same, are provided, wherein said non-human animals comprise a humanization of an endogenous cluster of differentiation (CD) gene, in particular a humanization of a CD47 gene. Said non-human animals may be described, in some embodiments, as having a genetic modification to an endogenous CD47 gene so that said non-human animals express a CD47 polypeptide that includes a human portion and a non-human portion (e.g., a murine portion).

1 . A rodent comprising a CD47 gene that comprises an endogenous rodent CD47 nucleotide sequence and a human CD47 nucleotide sequence, wherein the CD47 gene is operably linked to an endogenous rodent CD47 promoter. 
     
     
         2 . The rodent of  claim 1 , wherein the endogenous rodent CD47 nucleotide sequence comprises exons 1 and the exons downstream of exon 7 of an endogenous rodent CD47 gene. 
     
     
         3 . (canceled) 
     
     
         4 . The rodent of  claim 1 , wherein the human CD47 nucleotide sequence encodes amino acids 19-127, 19-141 or 16-292 of a human CD47 polypeptide. 
     
     
         5 . The rodent of  claim 1 , wherein the human CD47 nucleotide sequence comprises exons 2-7 of a human CD47 gene. 
     
     
         6 .- 12 . (canceled) 
     
     
         13 . The rodent of  claim 1 , wherein the CD47 gene encodes a CD47 polypeptide comprising an extracellular portion of a human CD47 polypeptide and an intracellular portion of an endogenous rodent CD47 polypeptide. 
     
     
         14 . The rodent of  claim 13 , wherein the CD47 polypeptide is translated in a cell of the rodent with a rodent signal peptide. 
     
     
         15 .- 29 . (canceled) 
     
     
         30 . The rodent of  claim 1 , wherein the rodent further comprises a SIRPα gene that encodes a SIRPα polypeptide comprising an extracellular portion of a human SIRPα polypeptide and an intracellular portion of an endogenous rodent SIRPα polypeptide. 
     
     
         31 .- 33 . (canceled) 
     
     
         34 . An isolated rodent cell or tissue whose genome comprises a CD47 gene that encodes a CD47 polypeptide comprising an extracellular portion of a human CD47 polypeptide and an intracellular portion of an endogenous rodent CD47 polypeptide. 
     
     
         35 . (canceled) 
     
     
         36 . The isolated cell or tissue of claim  31 , wherein the CD47 gene comprises exons 2-7 of a human CD47 gene and is operably linked to an endogenous rodent CD47 promoter. 
     
     
         37 . The isolated rodent cell or tissue of  claim 34 , wherein the cell or tissue further comprises a SIRPα gene that encodes a SIRPα polypeptide comprising an extracellular portion of a human SIRPα polypeptide and an intracellular portion of an endogenous rodent SIRPα polypeptide. 
     
     
         38 . (canceled) 
     
     
         39 . A rodent embryonic stem cell whose genome comprises a CD47 gene that encodes a CD47 polypeptide comprising an extracellular portion of a human CD47 polypeptide and an intracellular portion of an endogenous rodent CD47 polypeptide. 
     
     
         40 . The rodent embryonic stem cell of  claim 39 , wherein the CD47 gene comprises exons 2-7 of a human CD47 gene and is operably linked to an endogenous rodent CD47 promoter. 
     
     
         41 .- 44 . (canceled) 
     
     
         45 . A rodent embryo generated from the rodent embryonic stem cell of  claim 39 . 
     
     
         46 . (canceled) 
     
     
         47 . A method of making a rodent that expresses a CD47 polypeptide from an endogenous rodent CD47 locus, wherein the CD47 polypeptide comprises a human sequence, the method comprising:
 (a) inserting a genomic fragment into an endogenous rodent CD47 locus in a rodent embryonic stem cell, said genomic fragment comprising a nucleotide sequence that encodes a human CD47 polypeptide in whole or in part, thereby forming a humanized CD47 gene;   (b) obtaining a rodent embryonic stem cell comprising the humanized CD47 gene of (a); and,   (c) creating a rodent using the rodent embryonic stem cell of (b).   
     
     
         48 . The method of  claim 47 , wherein the human sequence comprises amino acids corresponding to residues 19-127, 19-141, 19-292 or 16-292 of a human CD47 polypeptide. 
     
     
         49 . The method of  claim 47 , wherein the nucleotide sequence that encodes a human CD47 polypeptide in whole or in part comprises exons 2-7 of a human CD47 gene. 
     
     
         50 . (canceled) 
     
     
         51 . The method of  claim 47 , wherein the humanized CD47 gene comprises exon 1 and the exons downstream of exon 7 of a rodent CD47 gene. 
     
     
         52 . The method of  claim 47 , wherein the method further comprises a step of inserting a genomic fragment into an endogenous rodent SIRPα locus of the rodent embryonic stem cell of (a), said genomic fragment comprising a nucleotide sequence that encodes a human SIRPα polypeptide in whole or in part. 
     
     
         53 . The method of  claim 52 , wherein the genomic fragment comprising a nucleotide sequence that encodes a human SIRPα polypeptide in whole or in part is inserted into an endogenous rodent SIRPα locus of the rodent embryonic stem cell of (a) prior to the insertion of a genomic fragment into the endogenous rodent CD47 locus. 
     
     
         54 . The method of  claim 47 , wherein the method further comprises breeding the rodent of (c) with a second rodent, said second rodent having a genome comprising a SIRPα gene that encodes a SIRPα polypeptide comprising an extracellular portion of a human SIRPα polypeptide and an intracellular portion of an endogenous rodent SIRPα polypeptide. 
     
     
         55 .- 108 . (canceled)