AAV-mediated expression using a synthetic promoter and enhancer
An isolated recombinant parvovirus vector comprising a synthetic enhancer comprising plurality of enhancer sequences operably linked to a promoter, and methods of using the vector, are provided.
1. An isolated recombinant parvovirus vector comprising a synthetic enhancer operably linked to a promoter, wherein the enhancer and promoter have a sequence having at least 90% nucleotide sequence identity to SEQ ID NO:27.
2. The recombinant parvovirus vector of claim 1 which is a bocavirus or an adeno-associated virus vector.
3. The recombinant parvovirus vector of claim 1 , wherein the promoter is operably linked to an open reading frame that encodes a prophylactic or therapeutic gene product.
4. The recombinant parvovirus vector of claim 1 , wherein the synthetic enhancer and promoter have at least 95%, 98%, 99% or more nucleic acid sequence identity to SEQ ID NO:27.
5. An isolated vector comprising a synthetic enhancer operably linked to a promoter, wherein the enhancer and promoter have a sequence having at least 90% nucleotide sequence identity to SEQ ID NO:27.
6. The vector of claim 5 , wherein the promoter is operably linked to an open reading frame.
7. The vector of claim 6 , wherein the open reading frame encodes a prophylactic or therapeutic gene product.
8. The vector of claim 5 , wherein the synthetic enhancer and promoter have at least 95%, 98%, 99% or more nucleic acid sequence identity to SEQ ID NO:27.
9. The vector of claim 5 which is a plasmid.
10. The recombinant parvovirus vector of claim 1 , wherein the enhancer and promoter have at least 95% nucleotide sequence identity to SEQ ID NO:27.
11. The recombinant parvovirus vector of claim 10 , wherein the enhancer and promoter have at least 98%, 99%, or more nucleic acid sequence identity to SEQ ID NO:27.
12. The recombinant parvovirus vector of claim 11 , wherein the enhancer and promoter comprise the sequence of SEQ ID NO:27.
13. The recombinant parvovirus vector of claim 3 , wherein the therapeutic gene product is cystic fibrosis transmembrane conductance regulator (CFTR) or CFTRΔR.
14. The recombinant parvovirus vector of claim 13 , wherein the CFTR or CFTRΔR is human CFTR or human CFTRΔR.
15. A recombinant adeno-associated vector comprising: (i) an enhancer operably linked to a promoter, wherein the enhancer and promoter comprise the sequence of SEQ ID NO:27; and (ii) a human CFTRΔR transgene operably linked to the promoter.