Aav vectors produced by insect cells comprising rep52 and rep78 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.
Claims25
An insect cell comprising a first nucleotide sequence coding for a first amino acid sequence and a second nucleotide sequence coding for a second amino acid sequence, wherein the first and second amino acid sequences comprise a common amino acid sequence of at least 100 amino acids with at least 90% amino acid identity between the first and second amino acid sequences, wherein the nucleotide sequences that encode the common amino acid sequence in the first and second amino acid sequences are less than 90% identical, and wherein the first nucleotide sequence encodes an amino acid sequence of a parvoviral Rep52 or 40 protein and the second nucleotide sequence encodes an amino acid sequence of a parvoviral Rep78 or 68 protein, and wherein the common amino acid sequences comprise the amino acid sequences from the second amino acid to the most C-terminal amino acid of the parvoviral Rep52 or 40 protein.
An insect cell according to any one of the preceding claims, wherein the nucleotide sequence coding for the common amino acid sequence in the nucleotide sequence with the improved codon usage bias comprises a continuous stretch of at least 25 codons all of which are common codons in accordance with Table 1 or Table 2.
An insect cell according to any one of the preceding claims, wherein the first and second nucleotide sequences are part of a nucleic acid construct wherein the first and second nucleotide sequence are each operably linked to expression control sequences for expression in an insect cell.
An insect cell according to any one of the preceding claims, wherein the parvoviral Rep proteins are adeno-associated virus (AAV) Rep proteins.
Dependencies are inferred from the claim text.