CD34-derived recombinant adeno-associated vectors for stem cell transduction and systemic therapeutic gene transfer
Novel adeno-associated virus (AAV) isolates in nucleotide and amino acid forms and uses thereof are provided. The isolates show tropism for certain target tissues, such as blood stem cells, liver, heart and joint tissue, and may be used to transduce stem cells for introduction of genes of interest into the target tissues. Discrete modified portions of the cap gene, VP1, VP2, and VP3, may be used alone or in combination in the present methods.
1. A recombinant nucleotide sequence which encodes an amino acid sequence of an AAV capsid or modified VP1, VP2 or VP3 portion thereof, wherein said amino acid sequence comprises a sequence selected from the group of SEQ ID NOS: 2-17.
2. The recombinant nucleotide sequence of claim 1 , wherein an AAV vector comprises the nucleotide sequence.
3. The recombinant nucleotide sequence of claim 2 , wherein the AAV vector is packaged, isolated and purified.
4. The recombinant nucleotide sequence of claim 1 , wherein the nucleic acid sequence comprises a sequence selected from the group of SEQ ID NOS: 20-35.
5. A method of transducing a cell comprising administering an AAV vector to the cell, wherein the AAV vector comprises
(i) a recombinant nucleotide sequence which encodes an amino acid sequence of an AAV capsid or modified VP1, VP2 or VP3 portion thereof, wherein said amino acid sequence comprises a sequence selected from the group of SEQ ID NOS: 2-17; and
(ii) a transgene.
6. The method of claim 5 , wherein the cell is a stem cell and the transgene is a reprogramming gene for the induction of induced pluripotent stem cells.
7. The method of claim 6 , wherein the stem cell is a hematopoietic stem cell (HSC) or a mesenchymal stem cell.
8. The method of claim 7 , wherein the HSC is a CD34 + HSC.
9. The method of claim 5 , wherein transducing a cell results in a transient transduction of the cell with the transgene.
10. The method of claim 9 , wherein the transgene is a zinc finger.
11. The method of claim 9 , wherein the transient transduction is performed in vitro and length of transduction time is controlled by culture conditions.
12. The method of claim 5 , wherein the cell is a liver cell.
13. The method of claim 5 , wherein the cell is a joint tissue cell.