ADAM6 mice
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.
1. A mouse whose genome comprises
(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 gene; 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 gene; and
(c) an ectopic nucleic acid sequence that encodes an Adam6 protein that is functional in the mouse,
so that the mouse is characterized in that:
(i) it is fertile; and
(ii) when it is immunized with an antigen, it generates antibodies comprising:
(A) heavy chain variable domains encoded by the one or more human V H , one or more human D H , and one or more human J H gene segments, linked to heavy chain constant domains encoded by the heavy chain constant region gene; and
(B) light chain variable domains encoded by the one or more human V L and one or more human J L gene segments linked to light chain constant domains encoded by the endogenous light chain constant region gene,
wherein the antibodies specifically bind the antigen.
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 mouse cell whose genome comprises:
(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.
18. The isolated cell of claim 17 , wherein the cell is an embryonic stem (ES) cell.
19. A method of making a mouse comprising introducing the ES cell of claim 18 into a host embryo gestating the host embryo in a surrogate mother, and allowing the surrogate mother to give birth to progeny derived in whole or in part from the ES cell.
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 .