Vectors with modified initiation codon for the translation of AAV-REP78 useful for production of AAV
First Claim
1. A first nucleic acid construct comprising a first nucleic acid sequence which encodes both Rep78 and Rep52 proteins from a Rep78 nucleotide sequence;
- the first nucleic acid sequence being operably linked to an expression control sequence that includes a promoter that is active in insect cells and is constructed such that both Rep78 and Rep52 proteins are produced upon expression in an insect cell.
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Abstract
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.
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Citations
21 Claims
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1. A first nucleic acid construct comprising a first nucleic acid sequence which encodes both Rep78 and Rep52 proteins from a Rep78 nucleotide sequence;
- the first nucleic acid sequence being operably linked to an expression control sequence that includes a promoter that is active in insect cells and is constructed such that both Rep78 and Rep52 proteins are produced upon expression in an insect cell.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
Specification