Genetically modified major histocompatibility complex mice

• US 10,219,493 B2
• Filed: 01/24/2017
• Issued: 03/05/2019
• Est. Priority Date: 10/28/2011
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Macdonald, Lynn, Murphy, Andrew J., Tu, Naxin, Gurer, Cagan, Voronina, Vera, Stevens, Sean
• Primary Examiner(s): Chen, Shin Lin
• Application Number:
  US15/413,785
• Publication Number:
  US 20170135329A1
• Time in Patent Office: 770 Days
• Field of Search: 800 18, 800 25, 5303873, 536 235, 536 241
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/70503 Immunoglobulin superfamily
• CPC Class Codes: C07K 14/70539 MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

The invention provides genetically modified non-human animals that express a humanized MHC II protein (humanized MHC II α and β polypeptides), as well as embryos, cells, and tissues comprising the humanized MHC II protein. Also provided are constructs for and methods of making said genetically modified non-human animals. Methods of using the genetically modified non-human animals to study various aspects of the human immune system are provided.

Claims

1. A mouse comprising at an endogenous Major Histocompatibility Complex II (MHC II) β

gene locus a nucleotide sequence encoding a chimeric human/mouse MHC II β

polypeptide that comprises a human portion and a mouse portion,

wherein the nucleotide sequence is expressed under regulatory control of an endogenous mouse MHC II β

promoter,

wherein the human portion of the chimeric human/mouse MHC II β

polypeptide comprises a human MHC II β

2 domain,

wherein the mouse portion of the chimeric human/mouse MHC II β

polypeptide comprises transmembrane and cytoplasmic domains of a mouse MHC II β

polypeptide, and

wherein the mouse expresses the chimeric human/mouse MHC II β

polypeptide on the surface of an antigen presenting cell of the mouse.

2. The mouse of claim 1, wherein the human portion of the chimeric MHC II β

polypeptide further comprises a human MHC II β

1 domain.

3. The mouse of claim 1, wherein the nucleotide sequence is expressed under regulatory control of endogenous mouse MHC II β

promoter and regulatory elements.

4. The mouse of claim 1, wherein the human portion of the chimeric human/mouse MHC II β

polypeptide is encoded by a human leukocyte antigen (HLA) class II gene selected from the group consisting of any β

chain gene of human HLA-DR, HLA-DQ, and HLA-DP.

5. The mouse of claim 4, wherein the human portion of the chimeric human/mouse MEW II β

polypeptide is encoded by a β

chain gene of human HLA-DR4.

6. A method of making the mouse of claim 1, comprising modifying an endogenous MHC II β

gene locus by replacing at the endogenous mouse MHC II β

gene locus a nucleotide sequence encoding a mouse MHC II β

2 domain with a nucleotide sequence encoding a human MHC II β

2 domain,

wherein the modified MHC II β

gene locus encodes, under regulatory control of an endogenous mouse MHC II β

promoter, a chimeric human/mouse MHC II β

polypeptide that comprises a human portion and a mouse portion,

wherein the human portion of the chimeric MHC II β

polypeptide comprises a human MHC II β

2 domain, and

wherein the mouse portion of the chimeric MHC II β

polypeptide comprises transmembrane and cytoplasmic domains of a mouse MHC II β

polypeptide.

7. The method of claim 6, wherein the human portion of the chimeric MHC II β

polypeptide is encoded by a human HLA class II gene selected from the group consisting of any β

chain gene of human HLA-DR, HLA-DQ, and HLA-DP.

8. The method of claim 6, wherein the mouse portion of the chimeric MHC II β

polypeptide is encoded by a β

chain gene of mouse H-2E, and the human portion of the chimeric MHC II β

polypeptide is encoded by a β

chain gene of human HLA-DR4.

9. The method of claim 6, wherein the replacement is made in a single ES cell, and the single ES cell is introduced into a mouse embryo to make a mouse.

10. The method of claim 6, further comprising replacing at the endogenous mouse MHC II β

gene locus a nucleotide sequence encoding a mouse MHC II β

1domain with a nucleotide sequence encoding a human MHC II β

1 domain.