Adeno-associated viruses engineered for selectable tropism
AssigneeRegents of the University of Minnesota
InventorDaniel Schmidt
Life of this patent
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Abstract
Methods to prepare recombinant adeno-associated virus (AAV) capsids with altered tropism and compositions having AAVs with altered tropism are provided.
Claims20
A method to prepare a plurality of vectors comprising DNA encoding a recombinant adeno-associated virus (AAV) VP2 capsid protein that comprises a peptide toxin that binds to a molecule of a eukaryotic cell, comprising:
- a) providing a DNA encoding at least one peptide toxin that binds to a eukaryotic cell, a DNA encoding at least one sorting signal, a DNA encoding at least one peptide linker, and a DNA encoding the VP2 of an AAV;
- and
- b) combining in a ligation reaction the DNA encoding the at least one peptide toxin, the DNA encoding the at least one sorting signal, the DNA encoding the at least one peptide linker, the DNA encoding the VP2 of the AAV, and a hast vector, thereby yielding a plurality of ligated DNA vectors having the host vector ligated to the DNA encoding the at least one peptide sorting signal ligated to the DNA encoding the at least one peptide toxin ligated to the DNA encoding the at least one peptide linker ligated to the DNA encoding the VP2, wherein the plurality of ligated DNA vectors encode an AAV VP2 having the peptide toxin that binds to the eukaryotic cell.
A composition comprising helper-virus free recombinant AAV(rAAV) comprising a chimeric capsid protein comprising a peptide toxin that binds to a molecule of a eukaryotic cell and VP2, and a rAAV genome, wherein the peptide toxin comprises PcTx1, MITx, Mambalgin-3, Mambalgin-1, APETx2, μ-CNTX-Pn1a, μO-CNTX MrVIB, μ-CNTX SIIIA, ProTx-I, Pterinotoxin-1, VSTX3, ω-CNTX MVIIA, DTX-K, αDTX, DTX-I, Hemitoxin, Charybdotoxin, κ-CNTX RIIIJ, TsTXK-α, GxTx, HaTx, VSTx1, CONK1, κ-PVIIA, α-CNTX PIA, α-BGTx, α-CNTX GID, α-CNTX PnIB, α-CNTX IMI, SLURP-1, or α-BGTx(V31 iso), or wherein the peptide toxin binds to a N-methyl-D-aspartate (NMDA) receptor.
A method to prepare helper free recombinant AAV the capsid of which comprises a peptide toxin that binds to a molecule of a eukaryotic cell, comprising:
- providing one or more vectors having a rAAV genome, nucleic acid encoding AAV VP1, AAY VP3, adenovirus E2A, adenovirus E4 and adenovirus VA and a host vector having an open reading frame comprising a DNA encoding at least one peptide sorting signal operably linked to a DNA encoding at least one peptide toxin which binds to the molecule of a eukaryotic cell operably linked to a DNA encoding the at least one peptide linker operably inked to a DNA encoding AAV VP2;
- and contacting a cell with the one or more vectors and the host vector in an amount that generates the helper free rAAV with a capsid having the peptide toxin.
Dependencies are inferred from the claim text.