Vectors with modified initiation codon for the translation of AAV-REP78 useful for production of AAV
First Claim
1. A nucleic acid construct comprising a nucleotide sequence encoding parvoviral proteins Rep78 and Rep52, wherein the nucleotide sequence consisting essentially of a single Rep78 coding sequence, a first expression control sequence, and a second expression control sequence, the single Rep78 coding sequence being operably linked to the first expression control sequence for expressing in the insect cell and the second expression control sequence located within the Rep78 coding sequence for expressing Rep52 in the 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 (AA V) 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
18 Claims
- 1. A nucleic acid construct comprising a nucleotide sequence encoding parvoviral proteins Rep78 and Rep52, wherein the nucleotide sequence consisting essentially of a single Rep78 coding sequence, a first expression control sequence, and a second expression control sequence, the single Rep78 coding sequence being operably linked to the first expression control sequence for expressing in the insect cell and the second expression control sequence located within the Rep78 coding sequence for expressing Rep52 in the insect cell.
Specification