ADAM6 MICE

• US 20130254911A1
• Filed: 05/09/2013
• Published: 09/26/2013
• Est. Priority Date: 02/25/2011
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: Regeneron Pharmaceuticals Incorporated
• Inventors: Macdonald, Lynn, Stevens, Sean, Murphy, Andrew J.
• Granted Patent: US 8,697,940 B2
• Time in Patent Office: Days
• US Class Current: 800/18
• CPC Class Codes: A01K 2207/15 Humanized animals
• 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 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: B60H 1/32281 comprising a single secondary circuit, e.g. at evaporator or condenser side
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 16/461 Igs containing Ig-regions, -domains or -residues form different species
• 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: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• 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

Mice are provided that comprise a reduction or deletion of ADAM6 activity from an endogenous ADAM6 locus, or that lack an endogenous locus encoding a mouse ADAM6 protein, wherein the mice comprise a sequence encoding an ADAM6 or ortholog or homolog or fragment thereof that is functional in a male mouse. In one embodiment, the sequence is an ectopic ADAM6 sequence or a sequence that confers upon a male mouse the ability to generate offspring by mating. Mice and cells with genetically modified immunoglobulin heavy chain loci that comprise an ectopic nucleotide sequence encoding a mouse ADAM6 or functional fragment or homolog or ortholog thereof are also provided.

Claims

1. A mouse comprising

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

and

(b) an insertion of one or more human V_L and one or more human J_L gene segments upstream of an endogenous light chain constant region; and

(c) an ectopic nucleic acid sequence that encodes an Adam6 protein that is functional in the mouse.

2. The mouse of claim 1, wherein the ectopic nucleic acid sequence is present at the endogenous heavy chain locus.

3. The mouse of claim 2, wherein the ectopic nucleic acid sequence is positioned within a human immunoglobulin heavy chain sequence.

4. The mouse of claim 3, wherein the ectopic nucleic acid sequence is positioned between two human V_H gene segments.

5. The mouse of claim 1, wherein the ectopic nucleic acid sequence is present at a position other than the endogenous heavy chain locus.

6. The mouse of claim 5, wherein the ectopic nucleic acid sequence is positioned between a V_H and a D_H gene segment.

7. The mouse of claim 1, wherein the insertion comprises at least 18 human V_H, at least one and up to 27 human D_H, and at least one and up to six human J_H gene segments.

8. The mouse of claim 1, wherein the insertion comprises at least 39 human V_H, at least one and up to 27 human D_H, and at least one and up to six human J_H gene segments.

9. The mouse of claim 1, wherein the insertion comprises 80 human V_H, at least one and up to 27 human D_H, and at least one and up to six human J_H gene segments.

10. The mouse of claim 1, wherein the mouse lacks endogenous V_H, D_H and J_H gene segments.

11. The mouse of claim 1, wherein the one or more human V_L and one or more human J_L gene segments are Vκ

and Jκ

gene segments.

12. The mouse of claim 11, wherein the endogenous light chain constant region is a mouse Cκ

region.

13. The mouse of claim 11, wherein the insertion comprises at least 16 human Vκ

and at least one and up to five human Jκ

gene segments.

14. The mouse of claim 11, wherein the insertion comprises at least 30 human Vκ

and at least one and up to five human Jκ

gene segments.

15. The mouse of claim 11, wherein the insertion comprises at least 40 human Vκ

and at least one and up to five human Jκ

gene segments.

16. The mouse of claim 11, wherein the mouse lacks endogenous Vκ

and Jκ

gene segments.

17. An isolated cell of the mouse of claim 1.

18. The isolated cell of claim 17, wherein the cell is an embryonic stem (ES) cell.

19. A method of making a mouse using the ES cell of claim 18.

20. The isolated cell of claim 17, wherein the cell is a B cell.

21. A hybridoma made from the B cell of claim 20.