Polynucleotides encoding rodent antibodies with human idiotypes and animals comprising same
The invention relates to polynucleotides, particularly chimeric polynucleotides useful for optimal production of functional immunoglobulins with human idiotypes in rodents. The invention further relates to rodents comprising such polynucleotides.
1. A chimeric polynucleotide comprising at least one human immunoglobulin (Ig) joining (J) region gene, an Ig constant (C) region gene, and a rat 3′ enhancer, wherein said rat 3′ enhancer is isolated from a 3′ region of a rat IgH locus, and comprises the sequence set forth as SEQ ID NO:1.
2. The chimeric polynucleotide as in claim 1 , further comprising at least one human Ig variable (V) region gene and/or a human Ig diversity (D) region gene.
3. The chimeric polynucleotide of claim 1 , wherein the C region gene is selected from the group consisting of a human C region gene and a rat C region gene.
4. The chimeric polynucleotide of claim 1 , wherein the C region gene comprises a rat C region gene.
5. The chimeric polynucleotide of claim 1 , wherein the C region gene comprises a constant region gene selected from the group consisting of Cμ and Cγ.
6. The chimeric polynucleotide of claim 1 , comprising a nucleic acid sequence substantially homologous to bacterial artificial chromosome (BAC) Annabel, or a portion thereof.
7. The chimeric polynucleotide of claim 2 , wherein the chimeric polynucleotide comprises a human Ig variable (V) region gene and said human Ig V region comprises at least one human V region gene isolatable from BAC6-VH3-11 and/or BAC3.
8. The chimeric polynucleotide of claim 1 , comprising in 5′ to 3′ order:
(a) a human Ig variable region comprising human V region genes in natural configuration and identical to human V region genes of BAC6-VH3-11 and/or BAC3; and
(b) a human Ig joining region comprising human J region genes in natural configuration and identical to human J region genes of the bacterial artificial chromosome (BAC) Annabel.
9. The chimeric polynucleotide of claim 2 , comprising nucleic acid sequences (a)-(e) in 5′ to 3′ order:
(a) the human Ig V region;
(b) the human Ig D region;
(c) the human Ig J region;
(d) the Ig C region; and
(e) the rat 3′ enhancer comprising SEQ ID NO:1.
10. The chimeric polynucleotide as in claim 9 , comprising a nucleic acid sequence substantially homologous to the nucleic acid sequence set forth as SEQ ID NO:2.
11. The chimeric polynucleotide as in claim 9 , comprising a nucleic acid sequence substantially homologous to the nucleic acid sequence set forth as SEQ ID NO: 11.
12. The chimeric polynucleotide as in any of claims 2 - 9 , wherein said V, D, and J regions are rearranged and form a complete exon encoding a heavy chain variable domain.
13. A rodent cell comprising the chimeric polynucleotide according to any one of claims 1 and 2 - 11 .
14. The rodent cell of claim 13 , further comprising a polynucleotide encoding a functional immunoglobulin comprising V, J, and C region genes and a nucleic acid sequence substantially homologous to a nucleic acid sequence selected from the group consisting of SEQ ID NO:6 and SEQ ID NO:7.
15. A rodent cell comprising the chimeric polynucleotide according to claim 12 .