Baculovirus expression systems
AssigneeGenethon
InventorsLionel Galibert, Otto-Wilhelm Merten, Monique Van Oers, Christel Riviere
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Abstract
The present invention relates to an optimized baculovirus construct useful for the production of virus(-like) particles or viral vectors, in particular viral vectors for gene therapy.
Claims9
A method for producing a recombinant adeno-associated virus (rAAV) vector, comprising infecting a cell with one or several recombinant baculovirus(es), each baculovirus comprising a recombinant baculovirus genome harboring a heterologous sequence;
- wherein the heterologous sequence from each of the one or several recombinant baculovirus(es) encodes a different part of said rAAV vector;
- and
- wherein each of the one or several recombinant baculovirus(es) comprises a recombinant baculovirus genome comprising the p26 and p74 baculoviral genes, and wherein the baculoviral genes cathepsin, chitinase and p10 are disrupted in each of the one or several recombinant baculovirus(es).
A method for producing an rAAV vector, comprising infecting an insect cell with three recombinant baculoviruses:
- (i) a baculovirus encoding the AAV Rep protein,
- (ii) a baculovirus encoding the AAV Cap protein;
- and
- (iii) a baculovirus encoding the AAV-ITR genome comprising a gene of interest between the two AAV-ITRs;
- wherein each of the recombinant baculoviruses comprises a recombinant baculovirus genome derived from the AcMNPV genome comprising the p26 and p74 baculoviral genes,
- wherein the baculoviral genes cathepsin, chitinase and p10 are disrupted in each of the recombinant baculoviruses, and
- wherein the p10 gene is disrupted without deleting the p10 promoter.
A method for producing an rAAV vector, comprising the step of infecting an insect cell with two recombinant baculoviruses:
- (i) a baculovirus encoding the AAV Rep protein and the AAV Cap protein;
- and
- (ii) a baculovirus encoding the AAV-ITR genome comprising a gene of interest between the two AAV-ITRs;
- wherein each of the recombinant baculoviruses comprises a recombinant baculovirus genome derived from the AcMNPV genome comprising the p26 and p74 baculoviral genes,
- wherein the baculoviral genes cathepsin, chitinase and p10 are disrupted in each of the recombinant baculoviruses, and
- wherein the p10 gene is disrupted without deleting the p10 promoter.
Dependencies are inferred from the claim text.