Recombinant aav vectors with altered immunogencity and methods of making the same
OwnerJohn Fraser Wrightinventor
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Abstract
The present invention provides methods of generating a recombinant AAV vector with reduced immunogenicity, comprising: providing eukaryotic cells with a nucleic acid comprising a sequence of interest that is flanked by AAV inverted terminal repeats, wherein the nucleic acid comprises CpG dinucleotide sites, wherein at least a portion of the CpG dinucleotide sites are methylated, wherein the eukaryotic cell expresses one or more other components necessary to achieve recombinant AAV biosynthesis, whereby the recombinant AAV vector is generated by the eukaryotic cell, wherein the generated recombinant AAV vector comprises nucleic acid wherein at least a portion of the CpG dinucleotide sites are methylated.
Claims25
A method of generating a recombinant AAV vector with reduced immunogenicity, comprising:
- providing eukaryotic cells with a nucleic acid comprising a sequence of interest that is flanked by AAV inverted terminal repeats,
- wherein the nucleic acid comprises CpG dinucleotide sites, wherein at least a portion of the CpG dinucleotide sites are methylated, wherein the eukaryotic cell expresses one or more other components necessary to achieve recombinant AAV biosynthesis,
- whereby the recombinant AAV vector is generated by the eukaryotic cell, wherein the generated recombinant AAV vector comprises nucleic acid wherein at least a portion of the CpG dinucleotide sites are methylated.
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A method of generating a recombinant AAV vector with increased immunogenicity, comprising:
- providing eukaryotic cells with a nucleic acid comprising a sequence of interest that is flanked by AAV inverted terminal repeats,
- wherein the nucleic acid has been engineered to be enriched in immunogenic CpG containing motifs,
- wherein the eukaryotic cell expresses one or more other components necessary to achieve recombinant AAV biosynthesis,
- wherein the eukaryotic cell optionally has a reduced capability of methylating CpG dinucleotide sites,
- whereby the recombinant AAV vector is generated by the eukaryotic cell, wherein the generated recombinant AAV vector comprises nucleic acid wherein at least a portion of the CpG dinucleotide sites are unmethylated.
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Dependencies are inferred from the claim text.