Viral vectors encoding recombinant FVIII variants with increased expression for gene therapy of hemophilia A
The present disclosure provides, among other aspects, codon-altered polynucleotides encoding Factor VIII variants for expression in mammalian cells. In some embodiments, the disclosure also provides mammalian gene therapy vectors and methods for treating hemophilia A.
1. A polynucleotide comprising the nucleotide sequence of SEQ ID NO: 13, wherein the polynucleotide encodes a Factor VIII polypeptide.
2. The polynucleotide of claim 1 , further comprising a promoter element operably linked to the polynucleotide encoding the Factor VIII polypeptide.
3. The polynucleotide of claim 2 , wherein the promoter element is a liver-specific promoter sequence upstream of the nucleotide sequence encoding the Factor VIII polypeptide.
4. The polynucleotide of claim 3 , further comprising an intron sequence positioned between the liver-specific promoter sequence and the nucleotide sequence encoding the Factor VIII polypeptide.
5. An adeno-associated virus (AAV) vector comprising a polynucleotide of claim 1 .
6. An adeno-associated virus (AAV) particle comprising a polynucleotide of claim 1 .
7. A host cell infected with an adeno-associated virus (AAV) particle comprising a polynucleotide of claim 1 .
8. A method for producing an adeno-associated virus (AAV) particle comprising introducing a polynucleotide of claim 1 into a mammalian host cell, wherein the polynucleotide is competent for replication in the mammalian host cell.
9. A method for treating hemophilia A comprising administering, to a patient in need thereof, an adeno-associated virus (AAV) particle according to claim 6 .
10. A method for transducing a host cell comprising contacting the host cell with an adeno-associated virus (AAV) particle according to claim 6 .