Factor IX gene therapy
The invention relates to a new, more potent, coagulation factor IX (FIX) expression cassette for gene therapy of haemophilia B (HB). Disclosed is a vector for expressing factor IX protein, the vector comprising a promoter, a nucleotide sequence encoding for a functional factor IX protein and an intron sequence, wherein the intron sequence is positioned between exon 1 and exon 2 of the nucleotide sequence encoding for a functional factor IX protein, and wherein the intron sequence has at least 80% identity to the sequence of SEQ ID NO. 1 as disclosed herein.
1. A vector for expressing a protein, the vector comprising:
a promoter operably linked to a polynucleotide encoding the protein;
wherein the promoter has at least 90% sequence identity to SEQ ID NO: 4.
2. The vector of claim 1 , wherein the promoter has at least 94% sequence identity to SEQ ID NO: 4.
3. The vector of claim 1 , wherein the promoter comprises SEQ ID NO: 4.
4. The vector of claim 1 , wherein the vector is an AAV vector.
5. The vector of claim 1 , wherein the vector is a single stranded vector.
6. The vector of claim 1 , wherein the promoter is 330-380 nucleotides in length.
7. The vector of claim 1 , wherein, the polynucleotide encoding a protein encodes a functional factor IX (FIX) protein.
8. The vector of claim 7 , wherein the polynucleotide encoding a functional factor IX (FIX) protein comprises an intron.
9. The vector of claim 8 , wherein the intron has at least 95% sequence identity to the sequence of SEQ ID NO: 1.
10. The vector of claim 8 , wherein the intron comprises SEQ ID NO: 1.
11. The vector of claim 8 , wherein the intron is positioned between exon 1 and exon 2 of the polynucleotide encoding a functional factor IX (FIX) protein.
12. The vector of claim 11 , wherein the polynucleotide encoding a functional FIX protein, including the intron between exon 1 and 2, has at least 80% sequence identity to SEQ ID NO: 3.
13. The vector of claim 11 , wherein the polynucleotide encoding a functional FIX protein, including the intron between exon 1 and 2, has at least 95% sequence identity to SEQ ID NO: 3.
14. The vector of claim 11 , wherein the polynucleotide encoding a functional FIX protein, including the intron between exon 1 and 2, comprises SEQ ID NO: 3.
15. The vector of claim 11 , wherein exon 1 is wildtype exon 1.
16. The vector of claim 8 , wherein the intron has at least 85% sequence identity to the sequence of SEQ ID NO: 1.
17. The vector of claim 8 , wherein the intron has at least 95% sequence identity to SEQ ID NO: 1, the coding region of the polynucleotide encoding the functional FIX protein has at least 95% sequence identity to SEQ ID NO: 2, and the promoter has at least 94% sequence identity to SEQ ID NO: 4, and wherein the vector is a single stranded vector.
18. The vector of claim 7 , wherein the coding regions of the polynucleotide encoding the functional FIX protein have at least 85% sequence identity to SEQ ID NO: 2.
19. The vector of claim 7 , wherein the coding regions of the polynucleotide encoding the functional FIX protein have at least sequence 95% identity to SEQ ID NO: 2.
20. The vector of claim 7 , wherein the coding regions of the polynucleotide encoding the functional FIX protein comprise SEQ ID NO: 2.