Genetic engineering of non-human animals for the production of chimeric antibodies
The invention provides non-human cells and mammals having a genome encoding chimeric antibodies and methods of producing transgenic cells and mammals. Certain aspects of the invention include chimeric antibodies, humanized antibodies, pharmaceutical compositions and kits. Certain aspects of the invention also relate to diagnostic and treatment methods using the antibodies of the invention.
1. A mouse whose genome comprises a transgene encoding a polypeptide comprising an immunoglobulin light chain variable region, wherein the transgene comprises (1) a plurality of immunoglobulin light chain variable (V) exons encoding canine immunoglobulin light chain variable (V) polypeptides; (2) non-coding sequences between the V exons; (3) a plurality of immunoglobulin light chain joining (J) coding sequences encoding canine immunoglobulin light chain joining (J) polypeptides; and (4) non-coding sequences between the J coding sequences; wherein the non-coding sequences between the V exons and the non-coding sequences between the J coding sequences are derived from mouse immunoglobulin light chain non-coding sequences and wherein the transgene is capable of undergoing gene arrangement and thereby upon expression to produce a polypeptide comprising the immunoglobulin light chain variable region.
2. The mouse according to claim 1 , wherein the non-coding sequences between the V exons and the non-coding sequences between the J coding sequences are selected from the group consisting of intronic sequences and cis regulatory sequences.
3. The mouse according to claim 2 , wherein the cis regulatory sequences are selected from promoters, enhancers, recombination signal sequences, splice acceptor sequences, and splice donor sequences.
4. The mouse according to claim 1 , wherein the V exons encode kappa light chain V(Vκ) polypeptides or lambda light chain V (Vλ) polypeptides.
5. The mouse according to claim 1 , wherein the V exons encode kappa light chain V (Vκ) polypeptides.
6. The mouse according to claim 1 , wherein the V exons encode lambda light chain V (Vλ) polypeptides.
7. The mouse according to claim 1 , wherein the transgene further comprises a coding sequence encoding a canine or mouse immunoglobulin light chain constant (C) polypeptide.
8. The mouse according to claim 7 , wherein the coding sequence encoding the immunoglobulin light chain C polypeptide encodes an immunoglobulin light chain constant lambda (Cλ) polypeptide or an immunoglobulin light chain constant kappa (Cκ) polypeptide.
9. The mouse according to claim 1 , wherein the transgene further comprises a coding sequence encoding a mouse immunoglobulin light chain constant (C) polypeptide.
10. The mouse according to claim 5 , wherein the transgene further comprises a coding sequence encoding a canine or mouse immunoglobulin light chain constant kappa (Cκ) polypeptide.
11. The mouse according to claim 6 , wherein the transgene further comprises a coding sequence encoding a canine or mouse immunoglobulin light chain constant lambda (Cλ) polypeptide.
12. The mouse according to claim 1 , wherein the transgene further comprises mouse non-coding sequences upstream of the V exons.
13. The mouse according to claim 12 , wherein the non-coding sequences upstream of the V exons are selected from promoters and enhancers.
14. The mouse according to claim 1 , wherein the transgene further comprises mouse non-coding sequences downstream of the J coding sequences.
15. The mouse according to claim 14 , wherein the non-coding sequences downstream of the J coding sequences are selected from polyadenylation sites and 3′ untranslated regions.
16. The mouse according to claim 11 , wherein the transgene further comprises a immunoglobulin light chain λ3′ enhancer.
17. The mouse according to claim 11 , wherein said transgene further comprises an immunoglobulin light chain 3′LCR, or a functional fragment thereof.
18. The mouse according to claim 17 , wherein said immunoglobulin light chain 3′LCR, or a functional fragment thereof, is from a mammal selected from the group consisting of human, non-primate and rat.
19. The mouse according to claim 1 , wherein the genome further comprises a second transgene encoding a canine immunoglobulin heavy chain, or a portion thereof.
20. A non-human mammalian cell whose genome comprises a transgene encoding a polypeptide comprising a canine immunoglobulin light chain variable region, wherein the transgene comprises (1) a plurality of immunoglobulin light chain variable (V) exons encoding canine immunoglobulin light chain variable (V) polypeptides; (2) non-coding sequences between the V exons; (3) a plurality of immunoglobulin light chain joining (J) coding sequences encoding canine immunoglobulin light chain joining (J) polypeptides; and (4) non-coding sequences between the J coding sequences; wherein the non-coding sequences between the V exons and the non-coding sequences between the J coding sequences are derived from mouse immunoglobulin light chain non-coding sequences and wherein the transgene is capable of undergoing gene arrangement and thereby upon expression to produce a polypeptide comprising the immunoglobulin light chain variable region.