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AssigneeuniQure IP B.V.
InventorsWilhelmus Theodorus Johannes Maria Christiaan Hermens, Saskia Jacoba Petronella Haast, Dennis Johan Biesmans, Andrew Christian Bakker
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The present invention relates nucleic acid constructs for the production of recombinant parvoviral (e.g. adeno-associated viral) vectors in insect cells, to insect cells comprising such constructs and to methods wherein the cells are used to produce recombinant parvoviral virions. The insect cells preferably comprise a first nucleotide sequence encoding the parvoviral rep proteins whereby the initiation codon for translation of the parvoviral Rep78 protein is a suboptimal initiation codon that effects partial exon skipping upon expression in insect cells. The insect cell further comprises a second nucleotide sequence comprising at least one parvoviral (AAV) inverted terminal repeat (ITR) nucleotide sequence and a third nucleotide sequence comprising a sequences coding for the parvoviral capsid proteins.
A method of producing a recombinant parvoviral virion in an insect cell comprising, transfecting an insect cell with a nucleotide sequence encoding parvoviral Rep proteins Rep78 and Rep52, the nucleotide sequence comprising a Rep78 coding sequence and a Rep52 coding sequence, the Rep78 coding sequence and the Rep52 coding sequence being operably linked to expression control sequences for expression of both a Rep78 protein and a Rep52 protein in an insect cell;
A method of producing a recombinant parvoviral virion in an insect cell comprising, transfecting an insect cell with a nucleic acid construct encoding parvoviral Rep proteins Rep78 and Rep52, the nucleic acid construct comprising:
Dependencies are inferred from the claim text.