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

Patent metadata

Field	Value
Publication number	US-10015953-B2
Application number	US-201615000751-A
Country	US
Kind code	B2
Filing date	Jan 19, 2016
Priority date	Dec 5, 2014
Publication date	Jul 10, 2018
Grant date	Jul 10, 2018

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).

First claim

We claim:

1. A genetically modified mouse whose genome comprises a replacement of a genomic fragment comprising exons 2-7 of a mouse CD47 gene at an endogenous mouse CD47 locus with a human genomic fragment comprising exons 2-7 of a human CD47 gene to form a humanized CD47 gene,
wherein the exons of said humanized CD47 gene consist of exon 1 of said mouse CD47 gene, exons 2-7 of said human CD47 gene, and the remaining exons downstream of exon 7 of said mouse CD47 gene,
wherein exons 2-7 of said human CD47 gene encode amino acids 16-292 of SEQ ID NO: 10,
wherein said humanized CD47 gene is under control of the endogenous mouse CD47 promoter at said endogenous mouse CD47 locus, and
wherein said mouse expresses a humanized CD47 protein encoded by said humanized CD47 gene.

2. The genetically modified mouse of claim 1 , whose genome further comprises a replacement of a genomic fragment comprising exons 2, 3 and 4 of a mouse SIRPα gene at an endogenous mouse SIRPα locus with a genomic fragment comprising exons 2, 3 and 4 of a human SIRPα gene to form a humanized SIRPα gene, wherein said humanized SIRPα gene is operably linked to the endogenous mouse SIRPα promoter at said endogenous mouse SIRPα locus, and wherein said humanized SIRPα gene encodes a humanized SIRPα protein comprising an extracellular portion of the human SIRPα protein encoded by said human SIRPα gene and an intracellular portion of the endogenous mouse SIRPα protein encoded by said mouse SIRPα gene.

3. An isolated mouse cell or tissue whose genome comprises a replacement of a genomic fragment comprising exons 2-7 of a mouse CD47 gene at an endogenous mouse CD47 locus with a human genomic fragment comprising exons 2-7 of a human CD47 gene to form a humanized CD47 gene,
wherein the exons of said humanized CD47 gene consist of exon 1 of said mouse CD47 gene, exons 2-7 of said human CD47 gene, and the remaining exons downstream of exon 7 of said mouse CD47 gene,
wherein exons 2-7 of said human CD47 gene encode amino acids 16-292 of SEQ ID NO: 10, and
wherein said humanized CD47 gene is under control of said endogenous mouse CD47 promoter at said endogenous mouse CD47 locus.

4. The isolated mouse cell or tissue of claim 3 , whose genome further comprises a replacement of a genomic fragment comprising exons 2, 3 and 4 of a mouse SIRPα gene at an endogenous mouse SIRPα locus with a genomic fragment comprising exons 2, 3 and 4 of a human SIRPα gene to form a humanized SIRPα gene, wherein said humanized SIRPα gene is operably linked to the endogenous mouse SIRPα promoter at said endogenous mouse SIRPα locus, and wherein said humanized SIRPα gene encodes a humanized SIRPα protein comprising an extracellular portion of the human SIRPα protein encoded by said human SIRPα gene and an intracellular portion of the mouse SIRPα protein encoded by said mouse SIRPα gene.

5. A mouse embryonic stem cell whose genome comprises a replacement of a genomic fragment comprising exons 2-7 of a mouse CD47 gene at an endogenous mouse CD47 locus with a human genomic fragment comprising exons 2-7 of a human CD47 gene to form a humanized CD47 gene,
wherein the exons of said humanized CD47 gene consist of exon 1 of said mouse CD47 gene, exons 2-7 of said human CD47 gene, and the remaining exons downstream of exon 7 of said mouse CD47 gene,
wherein exons 2-7 of said human CD47 gene encode amino acids 16-292 of SEQ ID NO: 10, and
wherein said humanized CD47 gene is under control of the endogenous mouse CD47 promoter at said endogenous mouse CD47 locus.

6. A mouse embryo comprising said mouse embryonic stem cell of claim 5 .

7. The genetically modified mouse of claim 2 , wherein said humanized SIRPα gene comprises exons 1, 5, 6, 7 and 8 of said mouse SIRPα gene.

Assignees

• Regeneron Pharma

Inventors

• Gurer Cagan
• Ioffe Ella
• Mujica Alexander
• Thurston Gavin

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A01K2227/105

  Murine · CPC title

• C12N15/8509

  for producing genetically modified animals, e.g. transgenic · CPC title

• A01K2217/072

  maintaining or altering function, i.e. knock in · CPC title

• A01K2267/0381

  Animal model for diseases of the hematopoietic system · CPC title