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 polynucleotide comprising a transgene encoding a polypeptide comprising an immunoglobulin heavy chain variable region, wherein the transgene comprises (1) a plurality of immunoglobulin heavy chain variable (V) exons encoding human immunoglobulin heavy chain V polypeptides; (2) non-coding sequences between the V exons; (3) a plurality of immunoglobulin heavy chain joining (J) coding sequences encoding human immunoglobulin heavy 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, rat, or a combination thereof immunoglobulin heavy chain non-coding sequences.
2. The polynucleotide of 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 polynucleotide of claim 2 , wherein the cis regulatory sequences are selected from the group consisting of promoters, enhancers, recombination signal sequences, splice acceptor sequences, and splice donor sequences.
4. The polynucleotide of claim 1 , wherein the transgene is capable of undergoing rearrangement to produce a polynucleotide sequence encoding a polypeptide comprising an immunoglobulin heavy chain variable region.
5. The polynucleotide of claim 1 , wherein the transgene further comprises one or more coding sequences encoding an immunoglobulin heavy chain constant (C) polypeptide.
6. An isolated non-human cell whose genome comprises the polynucleotide of claim 1 .
7. The polynucleotide according to claim 5 , wherein the one or more coding sequences encoding the immunoglobulin heavy chain C polypeptide encode mouse heavy chain C polypeptides.
8. The polynucleotide according to claim 1 , wherein the transgene further comprises mouse, rat, or a combination thereof non-coding sequences upstream of the V exons.
9. The polynucleotide according to claim 1 , wherein the transgene further comprises mouse, rat, or a combination thereof non-coding sequences downstream of the J coding sequences.
10. The polynucleotide according to claim 1 , wherein the transgene further comprises a plurality of immunoglobulin heavy chain diversity (D) coding sequences encoding immunoglobulin heavy chain D polypeptides.
11. The polynucleotide according to claim 10 , wherein the plurality of immunoglobulin heavy chain diversity (D) coding sequences encode human immunoglobulin heavy chain D polypeptides.
12. The polynucleotide according to claim 10 , wherein the transgene further comprises immunoglobulin heavy chain non-coding sequences between the D coding sequences, wherein the non-coding sequences between the D coding sequences are derived from mouse, rat or a combination thereof.
13. The polynucleotide according to claim 12 , wherein the non-coding sequences between the V exons, the non-coding sequences between the D coding sequences and the non-coding sequences between the J coding sequences are mouse immunoglobulin heavy chain non-coding sequences.
14. The polynucleotide according to claim 12 , wherein the non-coding sequences between the V exons, the non-coding sequences between the D coding sequences and the non-coding sequences between the J coding sequences are rat immunoglobulin heavy chain non-coding sequences.
15. The polynucleotide of claim 1 , wherein the transgene encoding a polypeptide comprising an immunoglobulin heavy chain variable region comprises (1) a plurality of immunoglobulin heavy chain variable (V) exons encoding human immunoglobulin heavy chain V polypeptides; (2) mouse immunoglobulin heavy chain non-coding sequences between the V exons; (3) a plurality of immunoglobulin heavy chain (D) coding sequences encoding human immunoglobulin heavy chain diversity D polypeptides; (4) mouse immunoglobulin heavy chain non-coding sequences between the D coding sequences; (5) a plurality of immunoglobulin heavy chain joining (J) coding sequences encoding human immunoglobulin heavy chain joining J polypeptides; (6) mouse immunoglobulin heavy chain non-coding sequences between the J coding sequences; and (7) one or more coding sequences encoding immunoglobulin mouse heavy chain constant (C) polypeptides.
16. The polynucleotide of claim 1 , wherein the transgene is a synthetic transgene.
17. The polynucleotide of claim 8 , wherein the non-coding sequences are selected from promoters and enhancers.
18. The polynucleotide of claim 9 , wherein the non-coding sequences are selected from polyadenylation sites and 3′ untranslated regions.