Humanized light chain mice

• US 9,706,759 B2
• Filed: 12/17/2012
• Issued: 07/18/2017
• Est. Priority Date: 12/20/2011
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Hosiawa, Karolina A., Stevens, Sean, Macdonald, Lynn, Murphy, Andrew J., Gurer, Cagan
• Primary Examiner(s): Ton, Thaian N
• Assistant Examiner(s): Montanari, David A
• Application Number:
  US13/716,238
• Publication Number:
  US 20130160153A1
• Time in Patent Office: 1,674 Days
• Field of Search: 800 18
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2217/15 Animals comprising multiple alterations of the genome, by transgenesis or homologous recombination, e.g. obtained by cross-breeding
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/462 Igs containing a variable region (Fv) from one specie and a constant region (Fc) from another
• CPC Class Codes: C12N 2800/204 of bacterial origin, e.g. BAC
• CPC Class Codes: C12N 2800/30 Vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT
• CPC Class Codes: C12N 9/6489 Metalloendopeptidases (3.4.24)

Abstract

Non-human animals, tissues, cells, and genetic material are provided that comprise a modification of an endogenous non-human heavy chain immunoglobulin sequence and that comprise an ADAM6 activity functional in a mouse, wherein the non-human animals express a human immunoglobulin heavy chain variable domain and a cognate human immunoglobulin λ light chain variable domain.

Claims

1. A transgenic mouse whose genome comprises:

(a) an insertion of one or more human Vλ

gene segments and one or more human Jλ

gene segments upstream of a mouse immunoglobulin light chain constant region gene,

(b) an insertion of one or more human V_H gene segments, one or more human D_H gene segments, and one or more human J_H gene segments upstream of a mouse immunoglobulin heavy chain constant region gene,

(c) an ectopic nucleotide sequence that encodes a mouse ADAM6 protein, wherein the mouse ADAM6 protein is expressed from the ectopic nucleotide sequence, and

(d) endogenous V_L gene segments and/or endogenous J_L gene segments that are incapable of rearranging to form an immunoglobulin light chain in the mouse.

2. The transgenic mouse of claim 1, wherein the mouse immunoglobulin light chain constant region gene is a mouse Cκ

or mouse Cλ

.

3. The transgenic mouse of claim 1, wherein the insertion of one or more human Vλ

gene segments and one or more human Jλ

gene segments comprises 12 to 40 human Vλ

gene segments and at least one human Jλ

gene segment.

4. The transgenic mouse of claim 1, wherein the one or more human Jλ

gene segments are selected from the group consisting of Jλ

1, Jλ

2, Jλ

3, Jλ

7, and a combination thereof.

5. The transgenic mouse of claim 1, wherein the one or more human Jλ

gene segments comprises at least four human Jλ

gene segments.

6. The transgenic mouse of claim 5, wherein the at least four human Jλ

gene segments comprise at least Jλ

1, Jλ

2, Jλ

3, and Jλ

7.

7. The transgenic mouse of claim 1, wherein the ectopic nucleotide sequence that encodes the mouse ADAM6 protein is present at the same location as compared to a wild-type type non-human ADAM6 locus.

8. The transgenic mouse of claim 1, wherein the ectopic nucleotide sequence that encodes the mouse ADAM6 protein is present between immunoglobulin gene segments.

9. The transgenic mouse of claim 8, wherein the immunoglobulin gene segments are heavy chain gene segments.

10. The transgenic mouse of claim 1, wherein all or substantially all endogenous Vκ

gene segments and endogenous Jκ

gene segments are replaced with one or more human Vλ

gene segments and one or more human Jλ

gene segments.

11. The transgenic mouse of claim 1, wherein all or substantially all endogenous Vλ

gene segments and endogenous Jλ

gene segments are replaced with one or more human Vλ

gene segments and one or more human Jλ

gene segments.

12. The transgenic mouse of claim 1, wherein the mouse further comprises a human Vκ

-Jκ

intergenic region from a human κ

light chain locus, wherein the human Vκ

-Jκ

intergenic region is contiguous with the one or more human Vλ

gene segments and one or more human Jλ

gene segments.

13. The transgenic mouse of claim 12, wherein the human Vκ

-Jκ

intergenic region is placed between a human Vλ

gene segment and a human Jλ

gene segment.

14. A cell derived from the transgenic mouse of claim 1.

15. The cell of claim 14, wherein the cell is a B cell.

16. The transgenic mouse of claim 1, wherein the mouse ADAM6 protein is ADAM6a or ADAM6b.