Back to results
US 2026/0144896 A1Published application
Modified cpg dinucleotides for recombinant viral vector production
AssigneeSiren Biotechnology, Inc.
InventorsNicole Kristen Paulk, Nathalie Andree Nancy Clement, Siti Nur Sarah Morris
Life of this patent
Estimating the term…
Abstract
The disclosure relates generally to methods of producing recombinant viral vectors with reduced immunogenicity.
Claims101
A nucleic acid, comprising:
- a helper polynucleotide comprising reduced CpG dinucleotides as compared to a parental equivalent.
A nucleic acid, comprising:
- a helper polynucleotide comprising increased methylation of CpG dinucleotides as compared to a parental equivalent.
A recombinant AAV vector comprising:
- a) a first nucleic acid comprising a nucleotide sequence having at least 90% sequence identity to the nucleic acid sequence of SEQ ID NOs:
- 13 or 14;
- and
- b) a second nucleic acid comprising a helper polynucleotide comprising a nucleotide sequence having at least about 90% sequence identity to the nucleic acid sequence of SEQ ID NOs:
- 10 or 11.
2 more elements
A recombinant AAV vector comprising:
- a) a first nucleic acid comprising a nucleotide sequence having at least 90% sequence identity to the nucleic acid sequence of SEQ ID NO:
- 14;
- and
- a second nucleic acid comprising one or more helper polynucleotides, the first helper polynucleotide of the one or more helper polynucleotides comprising a nucleotide sequence having at least about 90% sequence identity to the nucleic acid sequence of SEQ ID NO:
- 10 and the second helper polynucleotide of the one or more helper polynucleotides comprising a nucleotide sequence having at least about 90% sequence identity to the nucleic acid sequence of SEQ ID NO:
- or 11.
3 more elements
A recombinant AAV vector comprising:
- a) a first nucleic acid comprising a payload of interest;
- and
- b) a second nucleic acid comprising a helper polynucleotide, wherein the second nucleic acid has a reduced number of CpG dinucleotides as compared to a parental equivalent.
A recombinant AAV vector comprising:
- a) a first nucleic acid comprising a payload of interest;
- and
- b) a second nucleic acid comprising a helper polynucleotide, wherein the second nucleic acid has increased methylation of CpG dinucleotides as compared to a parental equivalent.
A composition comprising:
- a) a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease fused to an RNA methyltransferase or a nucleic acid encoding the CRISPR-associated endonuclease fused to an RNA methyltransferase;
- and
- b) one or more guide RNAs or a nucleic acid encoding the one or more guide RNAs, the one or more guide RNAs targeting a sequence for methylating a cytosine in a CpG dinucleotide.
A composition comprising:
- a) a nuclease fused to a methyltransferase;
- and
- b) a nucleic acid targeting a sequence comprising at least 97% sequence identity to any one of the nucleic acid sequences of SEQ ID NOs:
- 2-6 or a reverse complement thereof.
1 more element
A method for generating a recombinant AAV vector, comprising:
- a) providing cells with a nucleic acid comprising a payload of interest;
- and
- b) culturing the cells of step a) in a growth medium comprising cytosine or a derivative thereof to generate the recombinant AAV vector comprising methylation of at least about 25% of CpG dinucleotides.
A pharmaceutical composition comprising:
- a) a recombinant AAV vector comprising a payload of interest;
- b) a plurality of helper polynucleotides comprising reduced CpG dinucleotides;
- and
- c) a pharmaceutically acceptable excipient.
1 more element
A pharmaceutical composition comprising:
- a) a recombinant AAV vector comprising a payload of interest;
- b) a plurality of helper polynucleotides comprising increased methylation of CpG dinucleotides;
- and
- c) a pharmaceutically acceptable excipient.
1 more element
A pharmaceutical composition having less than about 50% CpG dinucleotides as compared to a parental equivalent, the pharmaceutical composition comprising
- a) a recombinant AAV vector comprising a payload of interest;
- b) a plurality of helper polynucleotides;
- and
- c) a pharmaceutically acceptable excipient,
- wherein the pharmaceutical composition comprises less than about 50% CpG dinucleotides.
2 more elements
Dependencies are inferred from the claim text.