Compositions and methods for the production of scAAV
AssigneeVoyager Therapeutics, Inc.
InventorsJames McLaughlin, Robert Kotin
Life of this patent
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Abstract
The present invention is directed to viral vectors and methods of their production and use.
Claims42
A parvoviral ITR nucleotide sequence comprising an engineered parvovirus Rep binding sequence region (eRBSR), wherein the eRBSR comprises one or more GCTC consensus motifs, wherein the eRBSR and parvoviral ITR are from the same parvoviral serotype, wherein, outside the eRBSR, the parvoviral ITR comprises a wild-type parvoviral ITR nucleotide sequence, wherein the eRBSR is not a wild-type JcDNV NS1 region, wherein the parvoviral ITR nucleotide sequence is capable of forming a double-stranded Rep binding sequence region, wherein the binding affinity between a Rep protein and the eRBSR is decreased relative to the binding affinity between the Rep protein and the wild-type parvoviral Rep binding sequence region, and wherein the Rep protein is selected from the group consisting of Rep DA-1, V8-865, AAV1 Rep, AAV3 Rep, AAV4 Rep, AAV6 Rep, AAV7 Rep, AAV8 Rep, AAV9 Rep, AAV10 Rep, or AAV12 Rep.
A parvoviral ITR nucleotide sequence comprising a Rep nicking sequence and an eRBSR;
- wherein the eRBSR is from 4 nucleotides to 20 nucleotides in length;
- wherein the eRBSR comprises one to four altered GCTC motif nucleotide sequences selected from the group consisting of N1CTC, N1NTC, N1CNC, N1CTN, GN2TC, GN2NC, GN2TN, GCN3C, GCN3N, and GCTN2, with N being an adenine, guanine, cytosine, thymine, or uracil, with Ni being an adenine, cytosine, thymine, or uracil, with N2 being an adenine, guanine, thymine, or uracil, and with N3 being an adenine, guanine or cytosine, wherein the parvoviral ITR and eRBSR are of the same parvoviral serotype, wherein, outside the eRBSR, the parvoviral ITR comprises a wild-type parvoviral ITR nucleotide sequence, wherein the eRBSR has decreased binding to a Rep protein compared to the wild-type parvoviral Rep binding sequence region, and wherein the Rep protein is selected from the group consisting of Rep DA-1, V8-865, AAV1 Rep, AAV3 Rep, AAV4 Rep, AAV6 Rep, AAV7 Rep, AAV8 Rep, AAV9 Rep, AAV10 Rep, or AAV12 Rep.
A parvoviral ITR nucleotide sequence comprising an eRBSR, wherein the eRBSR comprises an altered position of at least one GCTC motif present in the eRBSR compared to a wild-type parvoviral Rep binding sequence region, wherein the eRBSR differs from the wild-type parvoviral Rep binding sequence region by at least one nucleotide, wherein the eRBSR and parvoviral ITR are from the same parvoviral serotype, and wherein the eRBSR has decreased binding to a Rep protein compared to the wild-type parvoviral Rep binding sequence region, and wherein the Rep protein is selected from the group consisting of Rep DA-1, V8-865, AAV1 Rep, AAV3 Rep, AAV4 Rep, AAV6 Rep, AAV7 Rep, AAV8 Rep, AAV9 Rep, AAV10 Rep, or AAV12 Rep.
A parvoviral ITR nucleotide sequence comprising a Rep nicking sequence and an eRBSR;
- wherein the eRBSR comprises one to five GCTC motif nucleotide sequences;
- wherein the eRBSR comprises one to four altered GCTC motif nucleotide sequences selected from the group consisting of N1CTC, N1NTC, N1CNC, N5CTN, GN2TC, GN4NC, GN2TN, GCN3C, GCN3N, and GCTN2, with N being an adenine, guanine, cytosine, thymine, or uracil, with Ni being an adenine, cytosine, thymine, or uracil, with N2 being an adenine, guanine, thymine, or uracil, with N3 being an adenine, guanine or cytosine, with N4 being guanine, thymine, or uracil, and with N5 being adenine, cytosine, or uracil;
- wherein the eRBSR differs from a reference wild-type parvoviral Rep binding sequence region by at least one nucleotide;
- wherein the eRBSR and parvoviral ITR are from the same parvoviral serotype;
- wherein the eRBSR has decreased binding to a Rep protein compared to the wild-type parvoviral Rep binding sequence region, and wherein the Rep protein is selected from the group consisting of Rep DA-1, V8-865, AAV1 Rep, AAV3 Rep, AAV4 Rep, AAV6 Rep, AAV7 Rep, AAV8 Rep, AAV9 Rep, AAV10 Rep, or AAV12 Rep.
Dependencies are inferred from the claim text.