Adeno-associated virus factor VIII vectors
The present disclosure provides methods of generating multiplexed genetically modified animals, for example, porcine endogenous retrovirus (PERV)-inactivated pigs. The disclosure also provides methods of improving the birth rate of multiplexed genetically modified animals. In some embodiments, the present closure is concerned with the generation and utilization of porcine cells in which porcine endogenous retroviral (PERV) elements have been inactivated. In sonic embodiments, the PERV-free or PERV-reduced porcine cells are cloned to produce porcine embryos. In some embodiments, the PERV-free or PERV-reduced embryos may be grown into adult swine from which organs and/or tissues may be extracted and used for such purposes as xenotransplantation into non-porcine animals such as humans.
1. An adeno-associated virus (AAV) Factor VIII (FVIII) vector comprising a nucleic acid comprising an inverted terminal repeat (ITR), a liver-specific transcription regulatory region, and a functionally active FVIII coding region, wherein the functionally active FVIII coding region comprises nucleotides 923-5296 of SEQ ID NO: 9.
2. A method of producing a recombinant adeno-associated virus (AAV) particle comprising
A) culturing a cell that has been transfected with a AAV vector of claim 1 ; and
B) recovering recombinant AAV particle from the supernatant of the transfected cell.
3. A viral particle comprising the viral vector of claim 1 .
4. An isolated cell comprising the viral vector of claim 1 .
5. A method of treating a patient suffering from hemophilia A comprising intravenously administering to the patient an effective amount of an AAV FVIII vector of claim 1 .