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US 8,697,417 B2Granted
Baculoviral vectors comprising repeated coding sequences with differential codon biases
AssigneeuniQure IP B.V.
InventorsAndrew Christian Bakker, Wilhelmus Theodorus Johannes Maria Christiaan Hermens
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Abstract
The present invention relates to production of proteins in insect cells whereby repeated coding sequences are used in baculoviral vectors. In particular the invention relates to the production of parvoviral vectors that may be used in gene therapy and to improvements in expression of the viral rep proteins that increase the productivity of parvoviral vectors.
Claims30
An insect cell comprising a baculoviral vector comprising:
- (i) a first nucleotide sequence encoding a first amino acid sequence of a parvoviral Rep52 protein and is selected from the group consisting of:
- (a) SEQ ID NO:1;
- (b) SEQ ID NO:5;
- (c) A sequence of at least 75% identity with SEQ ID NO:2;
- (d) A sequence of at least 60% identity with SEQ ID NO:3;
- (e) A sequence of at least 70% identity with SEQ ID NO:4;
- (f) A sequence of at least 85% identity with SEQ ID NO:10;
- (g) A nucleotide sequence complementary to the full length sequence of any of (a)-(f);
- and
- (h) A nucleotide sequence that differs from the sequence of any of (c)-(f) due to degeneracy of the genetic code;
- and
- (ii) a second nucleotide sequence encoding a second amino acid sequence of a parvoviral Rep78 protein or a Rep68 protein,
- wherein the first and the second amino acid sequences comprise a homologous region of at least 100 amino acid residues with at least 90% identity beginning at the second residue and ending at the C-terminal residue of the parvoviral Rep 52, and
- wherein the portion of the first nucleotide sequence that encodes the homologous region and the portion of the second nucleotide sequence that encodes the homologous region are less than 90% identical.
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Dependencies are inferred from the claim text.