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US 9,580,691 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.
Claims22
An insect cell comprising a baculoviral vector comprising:
- a first nucleotide sequence encoding a first amino acid sequence of a parvoviral Rep52 protein selected from the group consisting of:
- (a) a nucleotide sequence of at least 85% identity with SEQ ID NO:
- 10;
- (b) a nucleotide sequence complementary to the full length sequence of (a);
- and
- (c) a nucleotide sequence that differs from the sequence of (a) due to degeneracy of the genetic code,
- and
- (ii) a second nucleotide sequence encoding a second amino acid sequence of a parvoviral Rep78 protein;
- wherein the first and the second amino acid sequences comprise a homologous region beginning at the second residue of the parvoviral Rep52 protein and ending at the C-terminal residue of the parvoviral Rep52 protein, the homologous region having at least 90% sequence identity between the first and the second amino acid sequences, and
- wherein the nucleotide sequences that encode the first and the second amino acid sequences of said region comprise a continuous stretch of at least 300 nucleotides that are less than 90% identical to one another.
7 more elements
An isolated nucleic acid construct comprising
- (i) a first nucleotide sequence encoding a first amino acid sequence of a parvoviral Rep52 protein selected from the group consisting of
- (a) a sequence that is SEQ ID NO:
- 10;
- (b) a nucleotide sequence complementary to the full length sequence of (a);
- and
- (c) a nucleotide sequence that differs from the sequence of (a) due to degeneracy of the genetic code;
- and
- (ii) a second nucleotide sequence encoding a second amino acid sequence of a parvoviral Rep78 protein;
- wherein the first and the second amino acid sequences comprise a homologous region beginning at the second residue of the parvoviral Rep52 protein and ending at the C-terminal residue of the parvoviral Rep52 protein, the homologous region having at least 90% sequence identity between the first and the second amino acid sequences, and
- wherein the nucleotide sequences that encode the first and the second amino acid sequences of said region comprise a continuous stretch of at least 300 nucleotides that are less than 90% identical to one another.
7 more elements
Dependencies are inferred from the claim text.