ADAM6 mice
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsLynn Macdonald, Sean Stevens, Andrew J. Murphy, Margaret Karow
Life of this patent
Estimating the term…
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.
Claims14
A method of determining a nucleotide sequence encoding a human heavy or light chain variable domain comprising the steps of:
- (a) immunizing a genetically modified mouse with an antigen, wherein the mouse:
- (i) has a genome that comprises:
- (A) one or more human VH gene segments, one or more human DH gene segments, and one or more human JH gene segments operably linked to a mouse heavy chain constant region at an endogenous mouse immunoglobulin heavy chain locus,
- (B) one or more human VL gene segments and one or more human JL gene segments operably linked to a light chain constant region, and
- (C) a nucleotide sequence that encodes a mouse ADAM6 protein, which nucleotide sequence is integrated in the genome of the mouse, wherein the mouse ADAM6 protein is expressed in the genetically modified mouse;
- (ii) generates antibodies when immunized with the antigen, wherein the antibodies each comprise a heavy chain including a human heavy chain variable domain operably linked to a mouse heavy chain constant domain and a light chain including a human light chain variable domain operably linked to a light chain constant domain;
- and
- (iii) is fertile;
- and
- (b) allowing the genetically modified mouse to mount an immune response to the antigen,
- (c) isolating a B cell from the genetically modified mouse that expresses an antibody that specifically binds the antigen, and
- (d) determining a human heavy or light chain variable region sequence that encodes a human heavy or light chain variable domain, respectively, of an antibody that specifically binds the antigen and that was generated by the genetically modified mouse.
Dependencies are inferred from the claim text.