Heterologous targeting peptide grafted AAVS
The disclosure in some aspects relates to recombinant adeno-associated viruses having distinct tissue targeting capabilities. In some aspects, the disclosure relates to gene transfer methods using the recombinant adeno-associated viruses. In some aspects, the disclosure relates to isolated AAV capsid proteins and isolated nucleic acids encoding the same.
1. A recombinant AAV (rAAV) comprising:
(a) a capsid protein having an N-terminally grafted heterologous targeting peptide, wherein:
(i) the capsid protein is a VP2 capsid protein that is not of an AAV2 serotype; and
(ii) the targeting peptide is a CNS-targeting polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 5, that is inserted between the first and second amino acid residues of the VP2 capsid protein; and
(b) a recombinant AAV vector comprising a transgene.
2. The rAAV of claim 1 , further comprising a linker conjugated to the C-terminus of the N-terminally grafted heterologous targeting peptide.
3. The rAAV of claim 2 , wherein the linker comprises at least one polypeptide repeat, each repeat comprising at least two glycine residues.
4. The rAAV of claim 3 , wherein the linker is of the formula [G] n S, wherein n is an integer in a range of 2 to 10.
5. The rAAV of claim 3 , wherein the linker comprises the formula [GGGGS] n , wherein n is an integer in a range of 1 to 4.
6. The rAAV of claim 5 , wherein the linker comprises SEQ ID NO: 28.
7. A recombinant AAV (rAAV) comprising:
(a) a capsid protein having an N-terminally grafted heterologous targeting peptide, wherein:
(i) the capsid protein is a VP2 capsid protein that is not of an AAV2 serotype; and
(ii) the targeting peptide is a CNS-targeting polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 7, that is inserted between the first and second amino acid residues of the VP2 capsid protein; and
(b) a recombinant AAV vector comprising a transgene.
8. The rAAV of claim 7 , further comprising a linker conjugated to the C-terminus of the N-terminally grafted heterologous targeting peptide.
9. The rAAV of claim 8 , wherein the linker comprises at least one polypeptide repeat, each repeat comprising at least two glycine residues.
10. The rAAV of claim 9 , wherein the linker is of the formula [G]S, wherein n is an integer in a range of 2 to 10.
11. The rAAV of claim 9 , wherein the linker comprises the formula [GGGGS] n , wherein n is an integer in a range of 1 to 4.
12. The rAAV of claim 11 , wherein the linker comprises SEQ ID NO: 28.
13. A recombinant AAV (rAAV) comprising:
(a) a capsid protein having an N-terminally grafted heterologous targeting peptide, wherein:
(i) the capsid protein is a VP2 capsid protein that is not of an AAV2 serotype; and
(ii) the targeting peptide is a CNS-targeting polypeptide encoded by a nucleic acid consisting of the nucleotide sequence set forth in SEQ ID NO: 29, that is inserted between the first and second amino acid residues of the VP2 capsid protein:
(b) a recombinant AAV vector comprising a transgene.
14. The rAAV of claim 13 , further comprising a linker conjugated to the C-terminus of the N-terminally grafted heterologous targeting peptide.
15. The rAAV of claim 14 , wherein the linker comprises at least one polypeptide repeat, each repeat comprising at least two glycine residues.
16. The rAAV of claim 15 , wherein the linker is of the formula [G]S, wherein n is an integer in a range of 2 to 10.
17. The rAAV of claim 15 , wherein the linker comprises the formula [GGGGS] n , wherein n is an integer in a range of 1 to 4, and wherein the linker optionally comprises SEQ ID NO: 28.