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US 11,279,949 B2Granted
Recombinant vectors comprising 2A peptide
AssigneeDenovo Biopharma LLC
InventorsDouglas J. Jolly, Amy H. Lin, Andrew Hofacre, Daniel J. Hogan, Derek G. Ostertag
Life of this patent
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Abstract
This disclosure provides modified recombinant retroviruses comprisings containing a 2A-peptide or peptide-like coding sequence operably linked to a heterologous polynucleotide. The disclosure further relates to cells and vector expressing or comprising such vectors and methods of using such modified vectors in the treatment of disease and disorders.
Claims22
A recombinant replication competent retrovirus comprising:
- a retroviral GAG protein;
- a retroviral POL protein;
- a retroviral envelope;
- a retroviral polynucleotide comprising Long-Terminal Repeat (LTR) sequences at the 3′ end of the retroviral polynucleotide sequence, a promoter sequence at the 5′ end of the retroviral polynucleotide, said promoter being suitable for expression in a mammalian cell, a gag nucleic acid domain, a pol nucleic acid domain and an env nucleic acid domain;
- a cassette comprising a 2A peptide or 2A peptide-like coding sequence that encodes the peptide of any one of SEQ ID Nos:
- 55-58 and is operably linked to a heterologous polynucleotide encoding a polypeptide having cytosine deaminase activity comprising the amino acid sequence as set forth in SEQ ID NO:
- 29, wherein the amino acid residue at position 10 and 152 of SEQ ID NO:
- 29 is F, D, M, L, or R, and wherein the cassette is positioned 5′ to the 3′ LTR and 3′ to the env nucleic acid domain encoding the retroviral envelope;
- and
- cis-acting sequences necessary for reverse transcription, packaging and integration in a target cell.
7 more elements
A recombinant replication competent retrovirus comprising:
- a retroviral GAG protein;
- a retroviral POL protein;
- a retroviral envelope;
- a retroviral polynucleotide comprising Long-Terminal Repeat (LTR) sequences at the 3′ end of the retroviral polynucleotide sequence, a promoter sequence at the 5′ end of the retroviral polynucleotide, said promoter being suitable for expression in a mammalian cell, a gag nucleic acid domain, a pol nucleic acid domain and an env nucleic acid domain;
- a cassette comprising regulatory domain operably linked to a first heterologous polynucleotide encoding a polypeptide having cytosine deaminase activity comprising the amino acid sequence as set forth in SEQ ID NO:
- 29, wherein the amino acid residue at position 10 and 152 of SEQ ID NO:
- 29 is F, D, M, L, or R, and operably linked to at least one 2A cassette comprising a GSG linker coding sequence and a 2A peptide or 2A peptide-like coding sequence that encodes a peptide of any one of SEQ ID Nos:
- 55-58 and is operably linked to a second heterologous polynucleotide, wherein the cassette is positioned 5′ to the 3′ LTR and 3′ to the env nucleic acid domain encoding the retroviral envelope and wherein the 2A cassette is downstream of the first heterologous polynucleotide;
- and
- cis-acting sequences necessary for reverse transcription, packaging and integration in a target cell.
7 more elements
Dependencies are inferred from the claim text.