VECTORS WITH MODIFIED INITIATION CODON FOR THE TRANSLATION OF AAV-REP78 USEFUL FOR PRODUCTION OF AAV

摘要

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.

主权项

1. A nucleic acid molecule comprising:
(a) a first nucleotide sequence that comprises a single open reading frame (ORF) encoding parvoviral Rep78 and Rep52 proteins, with a translation initiation site of the Rep78 protein being a suboptimal initiation codon selected from the group consisting of ACG, TTG, CTG and GTG; and (b) an expression control sequence that comprises nonanucleotide sequence SEQ ID NO:7 upstream of the translation initiation site operably linked to the single ORF.